CN115698719A

CN115698719A - Methods and compositions for cancer immunotherapy

Info

Publication number: CN115698719A
Application number: CN202180041949.4A
Authority: CN
Inventors: S·穆勒; B·Y·纳贝特; N·S·帕蒂尔; R·F·班奇罗
Original assignee: Genentech Inc
Current assignee: Genentech Inc
Priority date: 2020-06-12
Filing date: 2021-06-11
Publication date: 2023-02-03
Also published as: WO2021252977A1; JP2023529206A; WO2021252977A9; EP4165415A1; US20230392210A1

Abstract

The present invention provides diagnostic methods, therapeutic methods and compositions for treating cancer. The compositions and methods described herein can be used, for example, to identify patients with cancer who may benefit from treatment with a PD-L1 axis binding antagonist and to treat such patients accordingly. Using the compositions and methods of the present disclosure, a patient, such as a human cancer patient, may be determined to be likely to benefit from treatment with a PD-L1 axis binding antagonist if the patient exhibits the presence or elevated expression levels of any of the biomarkers disclosed herein. Exemplary PD-L1 axis binding antagonists that can be used in conjunction with the compositions and methods of the present disclosure are PD-L1 binding antagonists, such as anti-PD-L1 antibodies and antigen-binding fragments thereof, including attentizumab; and PD-1 binding antagonists, such as anti-PD-1 antibodies and antigen-binding fragments thereof.

Description

Methods and compositions for cancer immunotherapy

Sequence listing

This application contains a sequence listing that has been submitted electronically in ASCII format and is incorporated by reference herein in its entirety. The ASCII copy was created at 11.6.2021, named 50474-209WO2_sequence_listing_6.11.21_ST25, and was 398,463 bytes in size.

Cross reference to related patent applications

This application claims priority to U.S. provisional patent application No. 63/038,559, filed on 12/6/2020, the contents of which are incorporated herein by reference in their entirety.

Technical Field

The present invention relates to diagnostic and therapeutic methods for treating cancer using PD-L1 axis binding antagonists. Related kits and compositions are also provided.

Background

Cancer remains one of the most fatal threats to human health. In the united states, cancer affects nearly 130 million new patients each year, second only to heart disease, the second leading cause of death, accounting for approximately one-fourth of all deaths. In addition, it is predicted that cancer may be the first leading cause of death beyond cardiovascular disease within 5 years. Solid tumors are the leading cause of these deaths.

Human studies with immune checkpoint inhibitors have demonstrated the promise of using the immune system to control and eradicate tumor growth. The programmed death 1 (PD-1) receptor and its ligand programmed death ligand 1 (PD-L1) are immune checkpoint proteins associated with suppression of immune system responses during chronic infections, pregnancy, allografts, autoimmune diseases, and cancer. PD-L1 modulates immune responses by binding to the inhibitory receptor PD-1, which is expressed on the surface of T cells, B cells and monocytes. PD-L1 also exerts negative regulation of T cell function by interacting with another receptor, B7-1. The formation of the PD-L1/PD-1 and PD-L1/B7-1 complex down-regulates T cell receptor signaling, leading to down-regulation of T cell activation and inhibition of anti-tumor immune activity.

Despite major advances in the medical treatment of certain cancers, the 5-year overall survival rate of all cancers has only increased by about 10% over the past 20 years. In particular, malignant solid tumors metastasize and grow in an uncontrolled manner, which makes timely discovery and treatment extremely difficult.

Despite the great advances in cancer therapy, improved diagnostic and therapeutic methods and cancer therapies are sought.

Disclosure of Invention

The present disclosure provides therapeutic and diagnostic methods and compositions for treating an individual having cancer, e.g., lung cancer (e.g., non-small cell lung cancer (NSCLC)), endometrial cancer, colon adenocarcinoma, renal cell carcinoma, bladder cancer (e.g., urothelial Cancer (UC)), renal cancer (e.g., renal Cell Carcinoma (RCC)), and breast cancer (e.g., triple Negative Breast Cancer (TNBC)).

In one aspect, the invention features a method of identifying an individual having cancer who is likely to benefit from treatment with a PD-L1 axis binding antagonist, the method comprising determining the expression levels of two or more of the genes CD79A, CD19, BANK1, jchanin, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein the individual is identified as an individual who is likely to benefit from treatment with a PD-L1 axis binding antagonist if the immune score expression levels of the two or more genes are greater than reference immune score expression levels of the two or more genes.

In another aspect, the invention features a method of selecting a therapy for an individual having cancer, the method comprising determining the expression levels of two or more of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein the individual is identified as an individual likely to benefit from treatment comprising a PD-L1 axis binding antagonist if the immune score expression level of the two or more genes is greater than a reference immune score expression level of the two or more genes.

In some aspects, the immune score expression level of the two or more genes in the sample is higher than the reference immune score expression level, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a method of treating a subject having cancer, the method including: (a) Determining the expression level of two or more of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein the determination of the immune score expression level of the two or more genes in the sample is higher than a reference immune score expression level of the two or more genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a method of treating cancer in an individual, wherein the individual has been determined to have in a sample from the individual an immune score expression level of two or more of the genes CD79A, CD19, BANK1, jchan, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 that is greater than a reference immune score expression level for the two or more genes, the method comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a PD-L1 axis binding antagonist for use in a method of treating an individual having cancer, the method comprising: (a) Determining the expression level of two or more of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein the determination of the immune score expression level of the two or more genes in the sample is higher than a reference immune score expression level of the two or more genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a PD-L1 axis binding antagonist for use in treating cancer in an individual, wherein the individual has been determined to have in a sample from the individual an immune score expression level of two or more of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 that is higher than a reference immune score expression level of the two or more genes.

In some aspects, the immune score reference expression level is an immune score expression level of two or more genes in a reference population.

In some aspects, the reference population is a population of individuals with cancer.

In some aspects, the population of individuals includes a first subset of individuals who have received treatment with a PD-L1 axis binding antagonist and a second subset of individuals who have received treatment with a therapy that does not contain a PD-L1 axis binding antagonist.

In some aspects, the reference immune score expression level visibly differentiates each of the first subset of individuals from the second subset of individuals based on a significant difference between the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist and the responsiveness of the individual to treatment with the therapy without the PD-L1 axis binding antagonist above the reference immune score expression level, wherein the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist is significantly improved relative to the responsiveness of the individual to treatment with the therapy without the PD-L1 axis binding antagonist.

In some aspects, the therapy without a PD-L1 axis binding antagonist comprises an anti-neoplastic agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, or a combination thereof.

In some aspects, the therapy without the PD-L1 axis binding antagonist comprises a chemotherapeutic agent.

In some aspects, the chemotherapeutic agent is docetaxel.

In some aspects, responsiveness to treatment comprises prolongation of OS, prolongation of Progression Free Survival (PFS), or an increase in the determined optimal overall response (BCOR).

In some aspects, responsiveness to treatment is an extension of Overall Survival (OS).

In some aspects, the reference immune score expression level is the median of the expression levels of each of the two or more genes in the reference population.

In some aspects, the genes include three or more of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB 1.

In some aspects, the genes include four or more of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB 1.

In some aspects, the genes include five or more of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB 1.

In some aspects, the genes include six or more of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB 1.

In some aspects, the genes include seven or more of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB 1.

In some aspects, the genes include CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1.

In another aspect, the invention features a method of identifying an individual having cancer who is likely to benefit from treatment with a PD-L1 axis binding antagonist, the method comprising determining the expression level of one or more of the genes CD79A, CD19, BANK1, jcha in, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein the individual is identified as an individual who is likely to benefit from treatment with the PD-L1 axis binding antagonist if the immune score expression level of the one or more genes is greater than a reference immune score expression level for the one or more genes, wherein the benefit comprises an increase in Overall Survival (OS) of the individual as compared to treatment without the PD-L1 axis binding antagonist.

In another aspect, the invention features a method of selecting a therapy for an individual having cancer, the method comprising determining the expression level of one or more of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein the individual is identified as an individual likely to benefit from treatment with a PD-L1 axis binding antagonist in the event that the immune score expression level of the one or more genes is greater than a reference immune score expression level for the one or more genes, wherein the benefit comprises a prolongation of OS in the individual as compared to treatment without the PD-L1 axis binding antagonist.

In some aspects, the immune score expression level of one or more genes in the sample is greater than the reference immune score expression level, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a method of treating a subject having cancer, the method including: (a) Determining the expression level of one or more of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein determining the immune score expression level of the one or more genes in the sample is greater than a reference immune score expression level for the one or more genes identifies the individual as a likely to benefit from treatment comprising a PD-L1 axis binding antagonist, wherein the benefit comprises an increase in OS in the individual as compared to treatment without the PD-L1 axis binding antagonist; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a method of treating cancer in an individual, wherein the individual has been determined to have an immune score expression level of one or more of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual that is greater than a reference immune score expression level for the one or more genes, the method comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist, wherein in the event the immune score expression level of the one or more genes is greater than the reference immune score expression level for the one or more genes, identifying the individual as likely to benefit from treatment with a PD-L1 axis binding antagonist, wherein the benefit comprises a prolonged OS in the individual as compared to treatment without the PD-L1 axis binding antagonist.

In another aspect, the invention features a PD-L1 axis binding antagonist for use in a method of treating an individual having cancer, the method comprising: (a) Determining the expression level of one or more of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein determining the immune score expression level of the one or more genes in the sample is greater than a reference immune score expression level for the one or more genes identifies the individual as a likely to benefit from treatment comprising a PD-L1 axis binding antagonist, wherein the benefit comprises an increase in OS in the individual as compared to treatment without the PD-L1 axis binding antagonist; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a PD-L1 axis binding antagonist for use in treating cancer in an individual, wherein the individual has been determined to have an immune score expression level of one or more of genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual that is higher than a reference immune score expression level of the one or more genes, wherein in the event that the immune score expression level of the one or more genes is higher than the reference immune score expression level of the one or more genes, the individual is identified as an individual who is likely to benefit from treatment with the PD-L1 axis binding antagonist, wherein the benefit comprises an extended OS in the individual as compared to treatment without the PD-L1 axis binding antagonist.

In some aspects, the level of immune score expression of one of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 is determined.

In some aspects, the level of immune score expression of CD79A is determined.

In some aspects, the immune score reference expression level is an immune score expression level of one or more genes in a reference population.

In some aspects, the reference expression level of the immune score is based on a significant difference between the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist and the responsiveness of the individual to treatment with therapy that does not contain the PD-L1 axis binding antagonist above the reference expression level, distinctly distinguishing each of the first subset of individuals from the second subset of individuals, wherein the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist is significantly improved relative to the responsiveness of the individual to treatment with therapy that does not contain the PD-L1 axis binding antagonist.

In some aspects, the therapy without a PD-L1 axis binding antagonist comprises an anti-tumor agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, or a combination thereof.

In some aspects, the chemotherapeutic agent is docetaxel.

In some aspects, responsiveness to treatment comprises a prolongation of OS, a prolongation of PFS, or an increase in BCOR.

In some aspects, the immune score reference expression level is the median of the expression levels of each of the one or more genes in the reference population.

In some aspects, the median expression level is the median of the mean Z scores of the expression levels of each of the two or more genes in the reference population.

In some aspects, the genes include two or more of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB 1.

In some aspects, the two or more genes include TNFRSF17 and IGJ.

In some aspects, the two genes consist of TNFRSF17 and IGJ.

In some aspects, the genes include four or more of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1.

In some aspects, the genes include five or more of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1.

In some aspects, the genes include six or more of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1.

In some aspects, the genes include seven or more of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1.

In another aspect, the invention features a method of identifying an individual having cancer who is likely to benefit from treatment with a PD-L1 axis binding antagonist, the method including determining the expression level of one or more of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from the individual, wherein the individual is identified as an individual who is likely to benefit from treatment with a PD-L1 axis binding antagonist if the immune score expression level of the one or more genes is higher than the reference immune score expression level of the one or more genes.

In another aspect, the invention features a method of selecting a therapy for an individual having cancer, the method comprising determining the expression level of one or more of the genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from the individual, wherein the individual is identified as an individual likely to benefit from treatment with a PD-L1 axis binding antagonist if the immune score expression level of the one or more genes is greater than a reference immune score expression level of the one or more genes.

In some embodiments of any of the foregoing two aspects, the immune score expression level of the one or more genes in the sample is greater than the reference immune score expression level, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In other aspects, the invention features a method of treating an individual having cancer, the method including:

(a) Determining in a sample from the individual the expression level of one or more of the genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5, wherein the immune score expression level of the one or more genes in the sample is determined to be higher than a reference immune score expression level of the one or more genes; and

(b) Administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a method of treating cancer in an individual, wherein the individual has been determined to have an immune score expression level of one or more of the genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from the individual that is higher than a reference immune score expression level for the one or more genes, comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In some embodiments, the immune score reference expression level is an immune score expression level of one or more genes in a reference population.

In some embodiments, the reference population is a population of individuals having cancer.

In some embodiments, the population of individuals includes a first subset of individuals who have received treatment with a PD-L1 axis binding antagonist and a second subset of individuals who have received treatment with a therapy that does not contain a PD-L1 axis binding antagonist.

In some embodiments, the reference immune score expression level visibly differentiates each of the first subset of individuals from the second subset of individuals based on a significant difference between the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist and the responsiveness of the individual to treatment with the therapy without the PD-L1 axis binding antagonist above the reference immune score expression level, wherein the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist is significantly improved relative to the responsiveness of the individual to treatment with the therapy without the PD-L1 axis binding antagonist.

In some embodiments, the therapy without the PD-L1 axis binding antagonist comprises an anti-neoplastic agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, or a combination thereof.

In some embodiments, the therapy without the PD-L1 axis binding antagonist comprises a chemotherapeutic agent.

In some embodiments, the chemotherapeutic agent is docetaxel.

In some embodiments, responsiveness to treatment comprises prolongation of OS, prolongation of PFS, or an increase in BCOR.

In some embodiments, responsiveness to treatment includes an extension of Overall Survival (OS).

In some embodiments, the reference immune score expression level is the median of the expression levels of each of the one or more genes in the reference population.

In some embodiments, the genes include two or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

In some embodiments, the genes include three or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

In some embodiments, the genes include four or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

In some embodiments, the genes include five or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

In some embodiments, the genes include six or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

In some embodiments, the genes include seven or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

In some embodiments, the genes include eight or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

In some embodiments, the genes include nine or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

In some embodiments, the genes include ten or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

In some embodiments, the genes comprise eleven or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, or IGLL5.

In some embodiments, the genes include twelve or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, or IGLL5.

In some embodiments, the genes include thirteen or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, or IGLL5.

In some embodiments, the genes include MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5.

In another aspect, the invention features a method of identifying an individual having cancer who is likely to benefit from treatment with a PD-L1 axis binding antagonist, the method including determining the expression level of one or more of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from the individual, wherein the benefit includes an increase in OS of the individual as compared to treatment without the PD-L1 axis binding antagonist, where the immune score expression level of the one or more genes is greater than a reference immune score expression level for the one or more genes.

In another aspect, the invention features a method of selecting a therapy for an individual having cancer, the method comprising determining the expression level of one or more of the genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from the individual, wherein in the event the immune score expression level of the one or more genes is greater than a reference immune score expression level for the one or more genes, identifying the individual as an individual likely to benefit from treatment with a PD-L1 axis binding antagonist, wherein the benefit comprises an extended OS in the individual as compared to treatment without the PD-L1 axis binding antagonist.

In another aspect, the invention features a method of treating a subject having cancer, the method including:

(a) Determining in a sample from the individual the expression level of one or more of the genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5, wherein determining the immune score expression level of the one or more genes in the sample is higher than a reference immune score expression level of the one or more genes, thereby identifying the individual as an individual who is likely to benefit from treatment comprising a PD-L1 axis binding antagonist, wherein the benefit comprises a prolongation of OS in the individual as compared to treatment without the PD-L1 axis binding antagonist; and

In another aspect, the invention features a method of treating cancer in an individual, wherein the individual has been determined to have an immune score expression level of one or more of the genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7 and IGLL5 in a sample from the individual that is higher than a reference immune score expression level of the one or more genes, the method comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist, wherein the benefit comprises identifying the individual as an individual likely to benefit from treatment with the PD-L1 axis binding antagonist, wherein the benefit comprises a prolonged OS in the individual compared to treatment without the PD-L1 axis binding antagonist.

In some embodiments, the reference population is a population of individuals with cancer.

In some embodiments, the reference expression level of the immune score is based on a significant difference between the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist and the responsiveness of the individual to treatment with therapy that does not contain the PD-L1 axis binding antagonist above the reference expression level, distinctly distinguishing each of the first subset of individuals from the second subset of individuals, wherein the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist is significantly improved relative to the responsiveness of the individual to treatment with therapy that does not contain the PD-L1 axis binding antagonist.

In some embodiments, the therapy without a PD-L1 axis binding antagonist comprises an anti-tumor agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, or a combination thereof.

In some embodiments, the chemotherapeutic agent is docetaxel.

In some embodiments, the immune score reference expression level is the median of the expression levels of each of the one or more genes in the reference population.

In some embodiments, the median expression level is the median of the mean Z scores for the expression levels of each of the one or more genes in the reference population.

In some embodiments, the genes include seven or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, or IGLL 5.

In some embodiments, the genes include eleven or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

In some embodiments, the genes include twelve or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5.

In some embodiments, the genes include thirteen or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5.

In another aspect, the invention features a method of identifying an individual having cancer who is likely to benefit from treatment with a composition comprising a PD-L1 axis binding antagonist, the method including determining the presence of a Tertiary Lymphoid Structure (TLS) in a tumor sample from the individual, wherein the presence of TLS in the tumor sample identifies the individual as an individual who is likely to benefit from treatment with a composition comprising a PD-L1 axis binding antagonist.

In another aspect, the invention features a method of selecting a therapy for an individual with cancer, the method comprising determining the presence of TLS in a tumor sample from the individual, wherein the presence of TLS in the tumor sample identifies the individual as an individual who is likely to benefit from treatment comprising a PD-L1 axis binding antagonist.

In some aspects, the presence of TLS is determined in a sample from the individual, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a method of treating a subject having cancer, the method including: (a) Determining the presence of TLS in a tumor sample from the individual; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a method of treating cancer in an individual, wherein the individual has been determined to have TLS present in a tumor sample from the individual, comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a PD-L1 axis binding antagonist for use in a method of treating an individual having cancer, the method comprising: (a) Determining the presence of TLS in a tumor sample from the individual; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a PD-L1 axis binding antagonist for use in treating cancer in an individual, wherein the individual is determined to have TLS present in a tumor sample from the individual.

In some aspects, the presence of TLS is determined by histological staining, immunohistochemistry (IHC), immunofluorescence, or gene expression analysis.

In some aspects, the histological stain comprises a hematoxylin and eosin (H & E) stain.

In some aspects, IHC or immunofluorescence comprises detecting CD62L, L-selectin, CD40, or CD8.

In some aspects, the CD62L or L-selectin is detected using a MECA-79 antibody.

In some aspects, the gene expression analysis comprises determining the expression level of a TLS gene signature in the sample.

In some aspects, the TLS gene signature comprises one or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13.

In some aspects, the gene comprises two or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13.

In another aspect, the invention features a method of identifying an individual having cancer who may benefit from treatment comprising a PD-L1 axis binding antagonist, the method comprising determining expression levels of two or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual, wherein the individual is identified as an individual who may benefit from treatment comprising a PD-L1 axis binding antagonist if the immune score expression levels of the two or more genes are greater than reference immune score expression levels of the two or more genes.

In another aspect, the invention features a method of selecting a therapy for an individual having cancer, the method comprising determining an expression level of two or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual, wherein the individual is identified as an individual likely to benefit from treatment comprising a PD-L1 axis binding antagonist if the immune score expression level of the two or more genes is greater than a reference immune score expression level of the two or more genes.

In some aspects, the immune score expression level of two or more genes in the sample is greater than the reference immune score expression level, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a method of treating a subject having cancer, the method including: (a) Determining an expression level of two or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual, wherein determining an immune score expression level of the two or more genes in the sample is higher than a reference immune score expression level of the two or more genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a method of treating cancer in an individual, wherein the individual has been determined to have an immune score expression level of two or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual that is higher than a reference immune score expression level of the two or more genes, the method comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a PD-L1 axis binding antagonist for use in a method of treating an individual having cancer, the method comprising: (a) Determining an expression level of two or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual, wherein determining an immune score expression level of the two or more genes in the sample is higher than a reference immune score expression level of the two or more genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a PD-L1 axis binding antagonist for use in treating cancer in an individual, wherein the individual has been determined to have an immune score expression level of two or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual that is higher than a reference immune score expression level of the two or more genes.

In some aspects, the reference immune score expression level is an immune score expression level of two or more genes in a reference population.

In some aspects, the reference immune score expression level clearly distinguishes each of the first subset of individuals from the second subset of individuals based on a significant difference between the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist and the responsiveness of the individual to treatment with the therapy that does not contain the PD-L1 axis binding antagonist above the reference expression level, wherein the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist is significantly improved relative to the responsiveness of the individual to treatment with the therapy that does not contain the PD-L1 axis binding antagonist.

In some aspects, the chemotherapeutic agent is docetaxel.

In some aspects, the gene comprises three or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13.

In some aspects, the gene comprises four or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13.

In some aspects, the gene comprises five or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13.

In some aspects, the gene comprises six or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13.

In some aspects, the gene comprises seven or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13.

In some aspects, the gene comprises eight or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13.

In some aspects, the genes include nine or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13.

In some aspects, the gene comprises ten or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13.

In some aspects, the gene comprises eleven or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13.

In some aspects, the gene comprises CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13.

In some aspects, the expression level is a nucleic acid expression level.

In some aspects, the nucleic acid expression level is an mRNA expression level.

In some aspects, mRNA expression levels are determined by RNA-seq, RT-qPCR, multiplex qPCR or RT-qPCR, microarray analysis, SAGE, massARRAY technology, FISH, or combinations thereof.

In some aspects, mRNA expression levels are detected using RNA-seq.

In some aspects, the expression level is a protein expression level.

In some aspects, the protein expression level is determined by IHC, immunofluorescence, mass spectrometry, flow cytometry, and western blot, or a combination thereof.

In some aspects, the expression level is detected in tumor cells, tumor infiltrating immune cells, stromal cells, paracancerous normal tissue (NAT) cells, or a combination thereof.

In another aspect, the invention features a method of identifying an individual having cancer who is likely to benefit from treatment with a PD-L1 axis binding antagonist, the method including determining the number of B cells in a tumor sample from the individual, wherein the individual is identified as an individual who is likely to benefit from treatment with a PD-L1 axis binding antagonist if the number of B cells in the tumor sample is greater than a reference number of B cells.

In another aspect, the invention features a method of selecting a therapy for an individual having cancer, the method including determining a number of B cells in a tumor sample from the individual, wherein the individual is identified as an individual likely to benefit from treatment with a PD-L1 axis binding antagonist if the number of B cells in the tumor sample is greater than a reference number of B cells.

In some aspects, the number of B cells in the sample is greater than the reference number, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a method of treating a subject having cancer, the method including: (a) Determining the number of B cells in a tumor sample from the individual; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a method of treating cancer in an individual, wherein the individual has been determined to have a number of B cells in a tumor sample from the individual that is greater than a reference number of B cells, the method comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a PD-L1 axis binding antagonist for use in a method of treating an individual having cancer, the method comprising: (a) Determining the number of B cells in a tumor sample from the individual; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a PD-L1 axis binding antagonist for use in treating cancer in an individual, wherein the individual has been determined to have a number of B cells in a tumor sample from the individual that is higher than a reference number of B cells.

In some aspects, B cells include CD79+ B cells, igG + B cells, and/or plasma cells.

In another aspect, the invention features a method of identifying an individual having cancer who is likely to benefit from treatment with a compound comprising a PD-L1 axis binding antagonist, the method including determining whether the individual has clonally expanded B cells in a tumor sample from the individual, wherein the clonally expanded B cells in the sample identify the individual as an individual who is likely to benefit from treatment with a compound comprising a PD-L1 axis binding antagonist.

In another aspect, the invention features a method of selecting a therapy for an individual with cancer, the method including determining whether the individual has clonally expanded B cells in a tumor sample from the individual, wherein the clonally expanded B cells in the sample identify the individual as an individual likely to benefit from treatment comprising a PD-L1 axis binding antagonist.

In some aspects, the tumor sample comprises clonally expanded B cells, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a method of treating a subject having cancer, the method including: (a) Determining that the individual has clonally expanded B cells in a tumor sample from the individual; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a method of treating cancer in an individual, wherein the individual has been determined to have clonally expanded B cells in a tumor sample from the individual, the method comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a PD-L1 axis binding antagonist for use in a method of treating an individual having cancer, the method comprising: (a) Determining that the individual has clonally expanded B cells in a tumor sample from the individual; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another aspect, the invention features a PD-L1 axis binding antagonist for use in treating cancer in an individual, wherein the individual has been determined to have clonally expanded B cells in a tumor sample from the individual.

In some aspects, the clonally expanded B cells are clonally expanded plasma cells.

In some aspects, clonally expanded B cells are detected by measuring the diversity of B Cell Receptor (BCR) gene lineages in a tumor sample.

In some aspects, where a Shannon Diversity Index (SDI) of a BCR gene lineage in a tumor sample from an individual is lower than a reference SDI, the individual is identified as an individual who is likely to benefit from treatment comprising a PD-L1 axis binding antagonist.

In some aspects, the sample is a tissue sample, a cell sample, a whole blood sample, a plasma sample, a serum sample, or a combination thereof.

In some aspects, the tissue sample is a tumor tissue sample.

In some aspects, the tumor sample is a tumor tissue sample.

In some aspects, the tumor tissue sample comprises tumor cells, tumor infiltrating immune cells, stromal cells, NAT cells, or combinations thereof.

In some aspects, the tumor tissue sample is a formalin-fixed and paraffin-embedded (FFPE) sample, an archived sample, a fresh sample, or a frozen sample.

In some aspects, the tumor tissue sample is an FFPE sample.

In some aspects, the cancer is lung cancer, kidney cancer, bladder cancer, breast cancer, colorectal cancer, ovarian cancer, pancreatic cancer, stomach cancer, esophageal cancer, mesothelioma, melanoma, head and neck cancer, thyroid cancer, sarcoma, prostate cancer, glioblastoma, cervical cancer, thymus cancer, leukemia, lymphoma, myeloma, mycosis fungoides, merkel cell carcinoma, or a hematological malignancy.

In some aspects, the cancer is lung cancer, renal cancer, bladder cancer, or breast cancer.

In some aspects, the lung cancer is non-small cell lung cancer (NSCLC).

In some aspects, the NSCLC is non-squamous NSCLC.

In some aspects, the NSCLC is squamous NSCLC.

In some aspects, the benefit comprises a prolongation of OS in the individual, a prolongation of PFS in the individual, and/or an improvement in BCOR in the individual as compared to treatment without the PD-L1 axis binding antagonist.

In some aspects, the benefit comprises a prolongation of OS in the individual as compared to treatment without the PD-L1 axis binding antagonist.

In some aspects, the PD-L1 axis binding antagonist is selected from the group consisting of a PD-L1 binding antagonist, a PD-1 binding antagonist, and a PD-L2 binding antagonist.

In some aspects, the PD-L1 axis binding antagonist is a PD-L1 binding antagonist.

In some aspects, the PD-L1 binding antagonist inhibits the binding of PD-L1 to one or more of its ligand binding partners.

In some aspects, the PD-L1 binding antagonist inhibits the binding of PD-L1 to PD-1.

In some aspects, the PD-L1 binding antagonist inhibits the binding of PD-L1 to B7-1.

In some aspects, the PD-L1 binding antagonist inhibits the binding of PD-L1 to both PD-1 and B7-1.

In some aspects, the PD-L1 binding antagonist is an antibody or antigen-binding fragment thereof.

In some aspects, the antibody is selected from the group consisting of: alemtuzumab, MDX-1105, MEDI4736 (dewaluzumab), and MSB0010718C (avizumab).

In some aspects, the antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

In some aspects, the antibody comprises: a heavy chain variable region comprising the amino acid sequence of SEQ ID NO 7; and a light chain variable region comprising the amino acid sequence of SEQ ID NO 8.

In some aspects, the PD-L1 axis binding antagonist is a PD-1 binding antagonist.

In some aspects, the PD-1 binding antagonist inhibits the binding of PD-1 to one or more of its ligand binding partners.

In some aspects, the PD-1 binding antagonist inhibits the binding of PD-1 to PD-L1.

In some aspects, the PD-1 binding antagonist inhibits the binding of PD-1 to PD-L2.

In some aspects, the PD-1 binding antagonist inhibits the binding of PD-1 to both PD-L1 and PD-L2.

In some aspects, the PD-1 binding antagonist is an antibody or antigen-binding fragment thereof.

In some aspects, the antibody is selected from the group consisting of: MDX-1106 (nivolumab), MK-3475 (pembrolizumab), MEDI-0680 (AMP-514), PDR001, REGN2810, and BGB-108.

In some aspects, the PD-1 binding antagonist is an Fc fusion protein.

In some aspects, the Fc fusion protein is AMP-224.

In some aspects, the subject has not been treated for the cancer.

In some aspects, the subject has not been administered a PD-L1 axis binding antagonist.

In some aspects, the cancer is NSCLC, and wherein the individual does not have EGFR or ALK genomic tumor aberrations.

In some aspects, the subject has been previously treated for the cancer.

In some aspects, the subject has been treated for the cancer by administering to the subject a platinum-containing chemotherapeutic agent, and wherein the subject is non-responsive to the chemotherapeutic agent.

In some aspects, the PD-L1 axis binding antagonist is administered as a monotherapy.

In some aspects, the method further comprises administering an effective amount of one or more additional therapeutic agents.

The one or more additional therapeutic agents include an anti-tumor agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, an immunomodulatory agent, or a combination thereof.

In some aspects, the individual is a human.

In another aspect, the invention features a kit that includes a PD-L1 axis binding antagonist and instructions for administering the PD-L1 axis binding antagonist to an individual who has been identified as an individual who is likely to benefit from treatment comprising the PD-L1 binding antagonist according to any one of the methods disclosed herein.

In another aspect, the invention features a kit that includes a PD-L1 axis binding antagonist and instructions for administering the PD-L1 axis binding antagonist to an individual for whom treatment comprising the PD-L1 binding antagonist has been selected according to any one of the methods disclosed herein.

In another aspect, the invention features a kit for identifying an individual having cancer who is likely to benefit from treatment with a PD-L1 axis binding antagonist, the kit including reagents for determining the expression levels of two or more of the genes CD79A, CD19, BANK1, jchan, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein the individual is identified as an individual who is likely to benefit from treatment with a PD-L1 axis binding antagonist if the immune score expression levels of the two or more genes are greater than reference immune score expression levels for the two or more genes.

In another aspect, the invention features a kit for identifying an individual having cancer who is likely to benefit from treatment with a PD-L1 axis binding antagonist, the kit including reagents for determining the expression level of one or more of genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein the individual is identified as an individual who is likely to benefit from treatment with a PD-L1 axis binding antagonist if the immune score expression level of the one or more genes is greater than the reference immune score expression level for the one or more genes, wherein the benefit includes an increase in OS in the individual as compared to treatment without the PD-L1 axis binding antagonist.

In another aspect, the invention features a kit for identifying an individual having cancer who is likely to benefit from treatment with a PD-L1 axis binding antagonist, the kit including reagents for determining the expression level of one or more of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from the individual, wherein the benefit comprises an extended OS in the individual as compared to treatment without the PD-L1 axis binding antagonist, where the immune score expression level of the one or more genes is greater than a reference immune score expression level of the one or more genes.

In another aspect, the invention features a PD-L1 axis binding antagonist for use in a method of treating an individual having cancer, the method comprising:

(a) Determining in a sample from the individual the expression level of one or more of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7 and IGLL5, wherein the determination of the immune score expression level of the one or more genes in the sample is higher than the reference immune score expression level of the one or more genes; and

In another aspect, the invention features a PD-L1 axis binding antagonist for use in treating cancer in an individual, wherein the individual has been determined to have an immune score expression level of one or more of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from the individual that is higher than a reference immune score expression level of the one or more genes.

In another aspect, the invention features a PD-L1 axis binding antagonist for use in treating cancer in an individual, wherein the individual has been determined to have an immune score expression level of one or more of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from the individual that is higher than a reference immune score expression level for the one or more genes, wherein identifying the individual as an individual likely to comprise treatment with the PD-L1 axis binding antagonist would benefit if the immune score expression level of the one or more genes is higher than the reference immune score expression level for the one or more genes, wherein the benefit comprises a prolongation of the individual's OS as compared to treatment without the PD-L1 axis binding antagonist.

In another aspect, the invention features a kit for identifying an individual having cancer who is likely to benefit from treatment with a binding antagonist containing PD-L1 axis, the kit including reagents for determining the presence of Tertiary Lymphoid Structure (TLS) in a tumor sample from the individual, wherein the presence of TLS in the tumor sample identifies the individual as likely to benefit from treatment with a binding antagonist containing PD-L1 axis.

In another aspect, the invention features a kit for identifying an individual having cancer who may benefit from treatment comprising a PD-L1 axis binding antagonist, the kit comprising reagents for determining the expression level of two or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual, wherein the individual is identified as an individual who may benefit from treatment comprising a PD-L1 axis binding antagonist if the immune score expression level of the two or more genes is higher than a reference immune score expression level of the two or more genes.

In another aspect, the invention features a kit for identifying an individual having cancer who is likely to benefit from treatment with a PD-L1 axis binding antagonist, the kit including reagents for determining the number of B cells in a tumor sample from the individual, wherein the individual is identified as an individual who is likely to benefit from treatment with a PD-L1 axis binding antagonist if the number of B cells in the tumor sample is greater than a reference number of B cells.

In another aspect, the invention features a kit for identifying an individual having cancer who is likely to benefit from treatment with a PD-L1 axis binding antagonist, the kit including reagents for determining whether the individual has clonally expanded B cells in a tumor sample from the individual, wherein the clonally expanded B cells in the sample identify the individual as an individual who is likely to benefit from treatment with a PD-L1 axis binding antagonist.

Drawings

Figures 1A to 1C show that B-cell gene signature and CD79A are associated with atuzumab-mediated survival benefit in the POPLAR 2 phase study. FIG. 1A: differential gene expression analysis, showing enrichment of B cell gene signatures when comparing patients with OS ≤ 6 months (n = 24) to OS ≥ 12 months (n = 43). Yellow circles indicate B cell gene transcripts, and red circles indicate T _eff A gene transcript. Kaplan Meier (KM) curves comparing the probability of survival (n = 194) for patients with enriched (fig. 1B) B cell gene profiles (greater than median docetaxel patients =11.07 months, less than median docetaxel patients =9.23 months, less than median alemtuzumab patients =8.44 months) and (fig. 1C) CD79A genes (greater than median docetaxel patients =12.45 months, less than median docetaxel patients =9.23 months, less than median alemtuzumab patients =8.44 months).

Figures 2A to 2D show that patients with high B-cell gene signatures correlated with atuzumab-induced tumor responses in multiple phase 3 studies. KM curve comparing survival probability of patients with B-cell-rich gene profiles (both high (greater than median) and low (less than median), as shown): in the OAK trial (fig. 2A) (n = 727), the median greater docetaxel group patients =12.39 months, the median less docetaxel group patients =8.8 months, the median greater attritumab group patients =18.04 months, the median less attritumab group patients =7.79 months); and in (fig. 2B) BIRCH trial (n = 591), greater than median attrituximab group patients =17.74, less than median attritumab group patients =14.09 months. FIG. 2C: comparison of BCORs of patients with enriched B cell gene profiles in the docetaxel and alemtuzumab groups in the patient population was classified as Complete Remission (CR), partial Remission (PR), progressive Disease (PD), stable Disease (SD) according to RECIST V1.1 classification criteria. The p-value for docetaxel group was 0.34 and for alemtuzumab group was 0.00094 (Kruskal-Wallis test). FIG. 2D: progression Free Survival (PFS) profile in OAK trial (n = 727) (median greater docetaxel =4.17 months, median less docetaxel =2.83 months, median greater alemtuzumab =2.99 months, median less alemtuzumab =1.64 months).

Figures 3A to 3E show that patients responding to altlizumab have B cells and TLS infiltrated into the tumor, which appears to be associated with survival benefit. FIG. 3A: immunofluorescence representative images of pre-treatment lung adenocarcinoma samples showed high B cell gene profiles and were classified as atuzumab responders: CD79A is shown in green, CD8 in red, and Ki67 in blue. Scale bar: 100mm. FIG. 3B: representative H & E staining of lung adenocarcinoma from patients who responded to atuzumab showed the presence of TLS, as indicated by the marker. Scale bar: 500mm. FIG. 3C: correlation of CD79a gene expression, comparing patient tissues showing the presence or absence of TLS (. About.. P <0.001, paired t-test). FIG. 3D: correlation of CD3D gene expression, patient tissues showing the presence or absence of TLS were compared (.: p <0.01, paired t-test). FIG. 3E: in the POPLAR assay, the overall survival (in months) of the docetaxel and alemtuzumab groups correlated with the presence/absence of TLS (: p <0.05, paired t-test) (n = 194).

FIGS. 4A to 4D show immunostaining of TLS. Representative images of IHC from lung adenocarcinoma tissue of patients exhibiting high B cell gene signature and classified as attrituzumab responders, showing (fig. 4A) hematoxylin and eosin (H & E) staining in the presence of TLS (as shown by white circles), (fig. 4B) panCK and CD8 (red) staining, (fig. 4C) PNAd (peripheral lymph node addressen) (fig. 4D) CD40 staining. The scale bar is 20 μm.

Fig. 5A to 5D are KM curves comparing the survival probability of patients with enriched TLS gene characteristics: (fig. 5A) in the POPLAR trial (n = 194), greater than median docetaxel patients =10.63 months, less than median docetaxel patients =9.9 months, greater than median alemtuzumab patients =15.47 months, less than median alemtuzumab patients =8.54 months; and (figure 5B) in the OAK trial (n = 727), the median greater docetaxel group patients =10.28 months, the median less docetaxel group patients =10.28 months, the median greater attritumab group patients =14.32 months, the median less attritumab group patients =11.76 months. KM curves comparing survival probability of patients with enriched germinal center gene characteristics: (fig. 5C) in the POPLAR trial (n = 194), patients with greater than median docetaxel =8.87 months, less than median docetaxel =10.05 months, less than median alemtuzumab =9.72 months; and (fig. 5D) in the OAK trial (n = 727), the median greater docetaxel group patients =11.43 months, the median less docetaxel group patients =8.9 months, the median greater attritumab group patients =16.26 months, the median less attritumab group patients =9.95 months.

Fig. 6A to 6E show that patients who obtained the alemtuzumab-mediated benefit are enriched for B cell lineages. FIG. 6A: KM curves comparing the probability of survival (n = 727) for patients with plasma B cell 2-gene signature (median docetaxel =11.07 months, median less docetaxel =9.53 months, median greater alemtuzumab =16.43 months, median less alemtuzumab =7.82 months). BCR sequencing was done from limited patient samples (as shown), where the diversity of clonality was shown using shannon index: (fig. 6B) control patients (n = 3); (fig. 6C) PR (red) and SD (blue) patients prior to atlizumab treatment (n = 8); (fig. 6D) PD patients (n = 3), and a summary graph showing the change in shannon index (fig. 6E) is displayed.

Fig. 7 shows the enrichment of genes in the POPLAR assay and provides a list of genes enriched by the atuzumab responders and their HR and p values.

Fig. 8A and 8B show the association of B cells with PD-L1 status. A graph quantifying the association of B cell gene signatures with (fig. 8A) immune cell PD-L1 levels, where IC0=0%, IC1=1% to 5%, IC2=5% to 49%, IC3=50% or more (fig. 8B) immunity and tumor cell PD-L1 were determined by SP142 PD-L1 assay (Wilcoxon paired analysis).

FIGS. 9A to 9F showTLS prevalence and its association are shown. FIG. 9A: histology-based distribution of TLS (containing germinal centers) and lymphatic aggregates (no germinal centers). FIG. 9B: distribution of TLS in biopsy and resection samples. TLS presence identified by IHC and RNA sequencing-based (FIG. 9C) B cells, (FIG. 9D) T _eff And (FIG. 9E) association of genetic signatures of TLS (Wilcoxon paired analysis). FIG. 9F: association of overall survival (in months) of docetaxel and attrituximab groups with presence/absence of TLS in OAK assay (. About.. P:. Sup.)<0.001, paired t-test).

Fig. 10A to 10D show the association of B cell and TLS gene signatures with other biomarkers. Graphs quantifying the association of B cell characteristics with (fig. 10A) Tumor Mutation Burden (TMB) and (fig. 10B) STK11 mutation status. Graph showing the association of TLS gene signature with (fig. 10C) TMB and (fig. 10D) STK11 mutation status (Wilcoxon pairwise analysis).

Fig. 11A to 11C show the association of survival benefit with B cell immunophenotype. KM curves comparing the probability of survival for patients enriched in (n = 727): (fig. 11A) initial B-cell gene signature, greater than median docetaxel =10.71 months, less than median docetaxel =9.9 months, greater than median alemtuzumab =13.47 months, less than median alemtuzumab =11.79 months; (figure 11B) memory B cell characteristics, greater than median docetaxel =12.39 months, less than median docetaxel =8.8 months, greater than median alemtuzumab =17.64 months, less than median alemtuzumab =8.9 months; and (fig. 11C) plasma B cells, median greater docetaxel patients =11.53 months, median less docetaxel patients =9.72 months, median greater alemtuzumab patients =15.49 months, and median less alemtuzumab patients =9.72 months.

Fig. 12A to 12E show the enrichment of IgG subtype plasma cells in the atezumab responder. FIG. 12A shows% IgG; FIG. 12B shows the ratio of IgG to IgM; figure 12C shows% IgM; FIG. 12D shows the relative amounts of IgG compared to total IgG and IgM content; and figure 12E shows% IgA. BCR sequencing of different Ig domains compiled from limited patient samples, patients were classified as PR (brown), SD (blue) and PD (red) patients according to RECIST v1.1 before and after atuzumab treatment (n = 17).

Fig. 13A to 13D show the association of survival benefit with B cell immunophenotype. KM curves, comparing survival probabilities for patients enriched for: (fig. 13A) in the OAK trial, enriched for T cell effector gene signature and CD79A, with greater than median docetaxel =12.97 months, less than median docetaxel =9.12 months, greater than median alemtuzumab =15.9 months, less than median alemtuzumab =9.48 months; (fig. 13B) in the POPLAR trial, enriched for T cell effector gene signature and CD79A, with greater than median docetaxel =9.63 months, less than median docetaxel =9.35 months, less than median alemtuzumab =8.54 months; (figure 13C) enriched for T cell effector gene signature in OAK trial, with greater than median docetaxel patients =11.1 months, less than median docetaxel patients =9.82 months, greater than median alemtuzumab patients =15.34 months, less than median alemtuzumab patients =10.12 months; and (fig. 13D) enriched for T cell effector gene signature in the POPLAR trial with greater than median docetaxel patients =9.72 months, less than median docetaxel patients =9.23 months, greater than median alemtuzumab patients =15.47 months, less than median alemtuzumab patients =9.72 months.

Fig. 14A to 14C show that the association of B cell characteristics with OS benefit is consistent in the major subgroups. In the major subgroup, B cell signature was highly correlated with atlizumab-mediated OS benefit: (FIG. 14A) squamous vs non-squamous, (FIG. 14B) biopsy vs resection, and (FIG. 14C) lung tumor vs lymph node metastasis.

Fig. 15A to 15G show that intratumoral B cells were associated with increased OS in NSCLC patients receiving treatment with astuzumab. (FIG. 15A) shows volcano plots of differentially expressed genes (FDR P <0.05, absolute logFC ≧ 0.5) between patients with <6 months of OS from OAK (n = 205) and patients with >12 months of OS (n = 205) after treatment with atuzumab. (fig. 15B) same as fig. 15A, in patients receiving treatment with docetaxel. (FIGS. 15C to 15F) Kaplan Meier (KM) curves comparing the survival probability of patients enriched with CD79A, CD19, IFNG and the IFN-inducible chemokine CXCL 10. Gene expression was classified as high (upper T3 triplet) or low/medium (T1 and T2 triplet). (FIG. 15G) representative immunofluorescence images of pre-treatment lung adenocarcinoma tumors from two patients who responded to atuzumab (left panel) and two patients who did not respond to atuzumab. (scale bar: 100 μm).

Fig. 16A to 16D show the identification of three B-cell subsets in NSCLC tumors. (FIG. 16A) left: UMAP dimensionality reduction of 20,362 cells (spots). The same UMAP is given in the upper right corner. Right lower: from metastatic lymph nodes (mLN), non-metastatic lymph nodes (nLN), normal adjacent lung tissue (nlong), tumor biopsy (Tbio), and tumor resection (T res) in each cluster. Bottom center: relative mean expression of markers shown in the clusters of figure 16A. (FIG. 16B) left: the cell fraction (row) for each patient of the cluster given in fig. 16A. Right side: absolute cell numbers per patient in clusters in fig. 16A. (FIG. 16C) Violin diagram showing expression of marker genes in the cluster of FIG. 16A. (FIG. 16D) UMAP from B cell subsets of six purchased fresh NSCLC tumor samples, analyzed by CyTOF, recapitulated the presence of follicular B cells (HLA-DR +, CD 38-), germinal Center (GC) B cells (HLA-DR +, CD38+ Ki67 +) and plasma cells (HLA-DR-, CD38+ +).

Figures 17A and 17B show B cell subpopulation characteristics in batch RNAseq spectra. (FIG. 17A) hierarchical clustering of three B-cell features identified in OAK from scRNA-seq data. (FIG. 17B) scatter plot showing correlation between plasma cell, germinal center B cell and follicular B cell characteristics. Pearson R values are reported.

Figures 18A to 18F show that plasma cell characteristics independently predict response to atuzumab. (FIGS. 18A to 18C) Kaplan-Meier curves for OS for each of the three characteristics, divided into T3 (upper triplet) and T1-T2 (lower/middle triplet). Log rank p values are reported. (FIG. 18D) heatmap, showing the results of a Cox proportional hazards model that examines the risk ratio within and between groups. Dots represent statistically significant HR (p < 0.05). (fig. 18E) forest plot showing the significance of three B cell features and the previously reported 8 gene T effector feature (tGE 8) in the univariate interaction model, taking into account the interaction between the feature score and the treatment groups. (fig. 18F) forest plot showing the significance of the four features shown in fig. 18E in the multivariate analysis of the alemtuzumab group (left panel) and the docetaxel group (right panel). In all models, the features were classified as T3 and T1-T2.

Figures 19A to 19C show that patients with TLS/LA + tumors showed improved OS for treatment with atuzumab. (fig. 19A) H & E staining showing tumors with tertiary lymphoid structures (TLS, left panel), only lymphoid aggregates (middle panel), or neither (right panel) in representative samples from POPLAR. (fig. 19B) bar graph depicting the proportion of tumors with TLS, lymph Aggregate (LA) only, or neither in each treatment group in POPLAR. (FIG. 19C) Kaplan-Meier curve representing OS for tumors with TLS or LA and tumors without both given by treatment group.

FIGS. 20A to 20C show plasma cell TLS/LA + enriched tumors. (FIG. 20A) hierarchical clustering of three B-cell subpopulation characteristics. Samples were ordered by TLS/LA status. (FIG. 20B) volcano plots depicting differentially expressed genes between tumors with TLS and/or LA and tumors without both. Genes from three B cell characteristics are highlighted. (FIG. 20C) Violin plot showing characteristic z-scores of plasma cells, germinal center B cells and follicular B cells, grouped by TLS/LA status. Mann-Whitney p values are reported.

Fig. 21A to 21F provide more information about the data shown in fig. 15A to 15G. (FIG. 21A) shows volcanograms of differentially expressed genes (FDR P <0.05, absolute logFC ≧ 0.5) between patients from OAK with POPLAR <6 months (n = 58) and patients with OS >12 months (n = 87) after treatment with atuzumab. (fig. 21B) same as fig. 21A, in patients receiving treatment with docetaxel. (FIGS. 21C to 21F) Kaplan Meier (KM) curves comparing the survival probability of patients enriched with CD79A, CD19, IFNG and the IFN-inducible chemokine CXCL 10. Gene expression was classified as high (upper T3 triplet) or low/medium (T1 and T2 triplet).

Fig. 22A and 22B provide more information about the data shown in fig. 16A through 16D. (FIG. 22A) expression of putative signature genes of follicular B cells, plasma cells and GC B cells in non-B cell scRNA-seq compartments. Candidate markers for global deconvolution are highlighted, indicating the reason for the removal of the signature genes due to the global high background. (FIG. 22B) UMAP projection of scRNA-seq expression in B cells, depicting follicular B cells, plasma cells and GC B cells, summarizing CyTOF results for follicular B cells (HLA-DR +, CD 38-), germinal Center (GC) B cells (HLA-DR +, CD38+ Ki67 +) and plasma cells (HLADR-, CD38+ +).

FIG. 23 is the Pearson correlation of B cell subset signature genes in all samples in OAK, as described in example 1 below.

Fig. 24A through 24E provide more information about fig. 18A through 18F. (FIG. 24A) dichotomized plasma cell feature scores for three-level stratification for best overall remission within each group as objective remission or persistent stable disease (SD with PFS > 6 months) versus progressive disease or non-persistent stable disease (SD with PFS <6 months). The p value was Fisher's exact test. (FIGS. 24B to 24D) Kaplan-Meier curves for OS for each of the three features, divided into T3 (upper third) and T1-T2 (lower/middle third). Log rank P values are reported. (FIG. 24E) Kaplan-Meier curves for OS characteristic of plasma cells, divided into T3 and T1-2 in TCGA 365LUAD/LUSC data. The risk ratio and associated P-value for the overall survival period, which is characteristic of plasma cells, are shown.

Detailed Description

The present disclosure provides diagnostic methods, therapeutic methods, and compositions for treating cancer, e.g., lung cancer (e.g., non-small cell lung cancer (NSCLC)), bladder cancer (e.g., urothelial Cancer (UC)), kidney cancer (e.g., renal Cell Carcinoma (RCC)), and breast cancer (e.g., triple Negative Breast Cancer (TNBC)).

The present disclosure is based, at least in part, on the following findings: one or more of the biomarkers disclosed herein, e.g., the presence and/or expression level of any of the genes listed in any one of tables 1 to 17, the presence and/or expression level of a B cell characteristic (e.g., a plasma B cell characteristic), the presence of a Tertiary Lymphoid Structure (TLS), the presence and/or expression level of a TLS characteristic, the presence and/or number of B cells and/or the presence and/or of clonally expanded B cells, can be used to identify and select individuals who may benefit from treatment with a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attritumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

For example, as described in example 1 below, the present disclosure demonstrates that elevated expression levels of CD79A and other B cell signature genes, including plasma B cell signature genes, correlate with improved Overall Survival (OS) in NSCLC patients receiving treatment with the anti-PD-L1 antibody, atuzumab. Similarly, the presence of Tertiary Lymphoid Structures (TLS) and elevated expression levels of TLS signature genes are also associated with improved OS in NSCLC patients receiving treatment with the anti-PD-L1 antibody atelizumab. Thus, the biomarkers disclosed herein can be used, for example, to identify individuals who may benefit from treatment with a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attritumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), to select an optimized cancer therapy for a patient, and to provide personalized treatment methods for patients who may benefit.

I. Definition of

The term "about" as used herein refers to the usual range of error for the corresponding value as readily known to those of skill in the art. References herein to "about" a value or parameter include (and describe) embodiments that refer to the value or parameter itself.

As used herein, "administering" means a method of administering to a subject a dose of a compound (e.g., a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as alemtuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) or composition (e.g., a pharmaceutical composition, such as a pharmaceutical composition comprising a PD-L1 axis binding antagonist compound and/or composition used in the methods described herein can be administered, for example, intravenously (e.g., by intravenous infusion), subcutaneously, intramuscularly, intradermally, transdermally, intraarterially, intraperitoneally, intralesionally, intracranially, intraprostatally, intrapleurally, intratracheally, intranasally, intravitreally, intravaginally, intrarectally, topically, intratumorally, subconjunctivally, intravesicularlly, intracapsularly, mucosally, intraperically, intraoccularly, intraocularly, orally, topically, by inhalation, by injection, by infusion, by continuous infusion, by direct bathing target cells, by a catheter, by lavage, in a variety of the composition, or the disease condition to be treated, and the method to be treated.

"affinity" refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). As used herein, unless otherwise specified, "binding affinity" refers to intrinsic binding affinity that reflects a 1. The affinity of a molecule X for its partner Y can generally be determined by the dissociation constant (K) _D ) And (4) showing. Affinity can be measured by conventional methods known in the art, including those described herein. Specific illustrative and exemplary embodiments for measuring binding affinity are described below.

An antibody that is "affinity matured" refers to an antibody that has one or more alterations in one or more hypervariable regions (HVRs) that result in an improvement in the affinity of the antibody for an antigen compared to a parent antibody that does not have such alterations.

"amplification" as used herein generally refers to the process of producing multiple copies of a desired sequence. "multicopy" means at least two copies. "copy" does not necessarily mean perfect sequence complementarity or identity to the template sequence. For example, the copies may include nucleotide analogs, such as deoxyinosine, intentional sequence alterations (e.g., sequence alterations introduced by primers comprising sequences that are hybridizable, but not complementary, to the template), and/or sequence errors that occur during amplification.

The term "antibody" is used herein in the broadest sense and includes a variety of antibody structures, including, but not limited to, monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, so long as they exhibit the desired antigen-binding activity.

An "antibody fragment" refers to a molecule other than an intact antibody that comprises a portion of an intact antibody and binds to an antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, fv, fab '-SH, F (ab') ₂ (ii) a A diabody; a linear antibody; single chain antibody molecules (e.g., scFv); and multispecific antibodies formed from antibody fragments.

An "antibody that binds to the same epitope" as a reference antibody refers to an antibody that blocks binding of the reference antibody to its antigen by 50% or more in a competition assay, whereas a reference antibody blocks binding of the antibody to its antigen by 50% or more in a competition assay. An exemplary competition assay is provided herein.

The terms "anti-PD-L1 antibody" and "antibody that binds to PD-L1" refer to an antibody that is capable of binding to PD-L1 with sufficient affinity such that the antibody is useful as a diagnostic and/or therapeutic agent targeting PD-L1. In one embodiment, the anti-PD-L1 antibody binds to an unrelated, non-PD-L1 protein to less than about 10% of the extent of binding of the antibody to PD-L1, as measured, for example, by a Radioimmunoassay (RIA). In certain embodiments, the anti-PD-L1 antibody binds to an epitope of PD-L1 that is conserved among PD-L1 from different species. In certain embodiments, the anti-PD-L1 antibody is atelizumab. PD-L1 (programmed death ligand 1) is also known in the art as "programmed cell death 1 ligand 1", "PDCD1LG1", "CD274", "B7-H", and "PDL1". An exemplary human PD-L1 is shown in UniProtKB/Swiss-Prot accession number Q9NZQ7.1.

The term "anti-cancer therapy" refers to a therapy for treating cancer (e.g., lung cancer (e.g., non-small cell lung cancer (NSCLC), including non-squamous NSCLC and squamous NSCLC), bladder cancer (e.g., urothelial Cancer (UC)), renal cancer (e.g., renal Cell Carcinoma (RCC)), or breast cancer (e.g., triple-negative breast cancer (TNBC)))And (4) treating. Examples of anti-cancer therapeutic agents include, but are not limited to, e.g., PD-L1 axis binding antagonists (e.g., PD-L1 binding antagonists (e.g., anti-PD-L1 antibodies, such as alemtuzumab) or PD-1 binding antagonists (e.g., anti-PD-1 antibodies)), chemotherapeutic agents, growth inhibitors, cytotoxic agents, agents used in radiotherapy, anti-angiogenic agents, apoptotic agents, anti-tubulin agents, and other agents for treating cancer (e.g., anti-CD 20 antibodies), platelet-derived growth factor inhibitors (e.g., GLEEVEC) ^TM (imatinib mesylate)), COX-2 inhibitors (e.g., celecoxib), interferons, cytokines, antagonists (e.g., neutralizing antibodies) that bind to one or more of the following targets: PDGFR-beta, bl gamma S, APRIL, BCMA receptor, TRAIL/Apo2, other biologically active and organic chemical reagents, and the like. Combinations thereof are also included in the present invention.

An "article of manufacture" or "kit" as used interchangeably herein refers to any article of manufacture (e.g., package or container) or kit comprising at least one agent, e.g., a drug for treating a disease or condition (e.g., cancer, e.g., lung cancer (e.g., NSCLC, including non-squamous NSCLC and squamous NSCLC), bladder cancer (e.g., UC), renal cancer (e.g., RCC), or breast cancer (e.g., TNBC)), or a probe (e.g., a nucleic acid probe or antibody) for specifically detecting a biomarker described herein. In certain embodiments, the article of manufacture or kit is marketed, distributed, or sold as a means for performing the methods described herein.

The phrase "based on" as used herein means that information about one or more biomarkers is used to inform information provided on treatment decisions, package inserts or marketing/promotional guidelines, etc.

The term "B cell" as used herein refers to a lymphocyte that matures within the bone marrow and includes, but is not limited to, naive B cells, memory B cells, or plasma B cells (also referred to as plasma cells or effector B cells). B cells are also known in the art as "B lymphocytes". Unlike other lymphocytes such as T cells or natural killer cells, B cells can express a B Cell Receptor (BCR) on their plasma membrane.

The "B cell receptor" or "BCR" is a transmembrane receptor complex located on the B cell plasma membrane. BCRs include membrane-bound immunoglobulin (mIgG) moieties (e.g., mIgA, mIgG, mIgM, or mIgD) and signal transduction moieties composed of CD79A/CD79B heterodimers (also referred to as Ig- α/Ig- β). Each member of the CD79A/CD79B heterodimer spans the plasma membrane and includes a cytoplasmic tail that includes an immunoreceptor tyrosine-based activation motif (ITAM).

A "blocking" antibody or "antagonist" antibody is an antibody that inhibits or reduces the biological activity of the antigen to which it binds. Preferred blocking or antagonist antibodies substantially or completely inhibit the biological activity of the antigen.

"binding domain" refers to a portion of a compound or molecule that specifically binds to a target epitope, antigen, ligand, or receptor. Binding domains include, but are not limited to, antibodies (e.g., monoclonal, polyclonal, recombinant, humanized, and chimeric), antibody fragments, or portions thereof (e.g., fab fragment, fab' ₂ scFv antibodies, SMIPs, domain antibodies, diabodies, minibodies, scFv-Fc, affibodies, nanobodies, and VH and/or VL domains of antibodies), receptors, ligands, aptamers, and other molecules with identified binding partners.

The term "biomarker" as used herein refers to an indicator that can be detected in a sample, such as a predictive, diagnostic, and/or prognostic indicator (e.g., any gene listed in any one of tables 1-17, such as one or more of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, MZB1, CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, CXCL13, DERL3, JSRP1, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, and IGLC 7). Biomarkers can be used as indicators of particular subtypes of a disease or disorder (e.g., cancer, such as lung cancer (e.g., NSCLC, including non-squamous NSCLC and squamous NSCLC), bladder cancer (e.g., UC), renal cancer (e.g., RCC), or breast cancer (e.g., TNBC)) characterized by particular molecular, pathological, histological, and/or clinical features. In some embodiments, the biomarker is a gene. Biomarkers include, but are not limited to, polynucleotides (e.g., DNA and/or RNA), polynucleotide copy number alterations (e.g., DNA copy number), polypeptides and polynucleotide modifications (e.g., post-translational modifications), carbohydrates, glycolipid-based molecular markers, cells (e.g., B cells), and/or histological structures (e.g., tertiary lymphoid structures).

The terms "biomarker signature," "biomarker expression signature," or "expression signature" are used interchangeably herein and refer to the level of immune expression of one or a combination of biomarkers (e.g., any of the genes listed in any one of tables 1-17 (e.g., CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, MZB1, CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, jscl 9, CXCL10, CXCL11, CXCL13, DERL3, crp 1, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, and IGLC 7)) whose expression is an indicator (e.g., predictive, diagnostic, and/or prognostic indicator). Biomarker signatures can be used as indicators of particular subtypes of a disease or disorder (e.g., cancer, such as lung cancer (e.g., NSCLC, including non-squamous NSCLC and squamous NSCLC), bladder cancer (e.g., UC), renal cancer (e.g., RCC), or breast cancer (e.g., TNBC)) characterized by particular molecular, pathological, histological, and/or clinical features. In some embodiments, the biomarker signature is a "genetic signature". The term "gene signature" is used interchangeably with "gene expression signature" and refers to one or a combination of polynucleotides whose expression is an indicator (e.g., a predictive, diagnostic, and/or prognostic indicator). The gene signature may be, for example: b cell gene signatures (e.g., one or more of genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and/or MZB 1), initial B cell gene signatures (e.g., one or more of genes ABCB4, BCL7A, BEND5, BRAF, IL4R, LINC00921, MEP1A, MICAL3, NIPSNAP3B, PSG2, SELL, TCL1A, UGT1A8, and/or ZNF 286A), memory B cell gene signatures (e.g., one or more of the genes AIM2, ALOX5, CLCA3P, FAM65B, IFNA10, IL7, NPIPB15, SP140, TNFRSF13B, TRAF4, and/or ZBTB 32), plasma cell gene signature (e.g., one or more of the genes DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and/or IGLL5, and/or one or more of the genes ABCB9, AMPD1, ANGPT4, ATXN8OS, C11, CCr10, HIST1H2AE, HIST1H2BG, IGHE, KCNA3, KCNG2, LOC100130100, MAN1A1, MANEA, MAST1, MROH7, MZB1, PAX7, PDK1, RASGRP3, REN, SPAG4, ST6GAL 4, TGM5, UGT2B17, ZBP1, and/or ZNF 16), a characteristic of a T effector (e.g., one or more of the genes CCL2, CCL3, CCL4, NAC 5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and/or CXCL 13) or a sub-characteristic of a T effector (e.g., one or more of the genes CD8A, TLMES, GZMA, TBX21, IFNG, CXCL9, CXCL11, and/or CXCL 10). In some embodiments, the biomarker signature is a "protein signature. The term "protein characteristic" is used interchangeably with "protein expression characteristic" and refers to one or a combination of polypeptides whose expression is an indicator (e.g., a predictive, diagnostic, and/or prognostic indicator).

The term "CD79A" as used herein, unless otherwise indicated, refers to a cluster of differentiated CD79A genes, including any native CD79A from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). CD79A is also known in the art as Ig- α, B cell antigen receptor complex associated protein α chain and MB-1 membrane glycoprotein. The term encompasses "full-length" unprocessed CD79A, as well as any form of CD79A resulting from processing in a cell. The term also encompasses naturally occurring variants of CD79A, such as splice variants or allelic variants. The nucleic acid sequence of exemplary human CD79A is set forth in SEQ ID NO 13 (NCBI reference: NM-001783.4). The amino acid sequence of an exemplary protein encoded by human CD79A is shown in SEQ ID NO 14 (UNIPROT) ^TM Accession number P11912-1).

The term "SLAMF7" as used herein, unless otherwise indicated, refers to any native SLAMF7 (signaling lymphocyte activating molecule (SLAM) family member) from any vertebrate source7) The vertebrate source includes mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length" unprocessed SLAMF7, as well as any form of SLAMF7 that results from processing in the cell. The term also encompasses naturally occurring variants of SLAMF7, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human SLAMF7 is set forth in SEQ ID NO:15 (NCBI reference sequence: NM-021181.5). The amino acid sequence of an exemplary protein encoded by human SLAMF7 is shown in SEQ ID NO 16 (UNIPROT) ^TM Accession number Q9NQ 25-1).

The term "BTK" as used herein, unless otherwise indicated, refers to any native BTK (Bruton tyrosine protein kinase) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length" unprocessed BTK, as well as any form of BTK that results from processing in a cell. The term also encompasses naturally occurring variants of BTK, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human BTK is set forth in SEQ ID NO:17 (NCBI reference sequence: NM-000061.2). The amino acid sequence of an exemplary protein encoded by human BTK is shown in SEQ ID NO 18 (UNIPROT) ^TM Accession number Q06187-1).

The term "TNFRSF17" as used herein, unless otherwise indicated, refers to any native TNFRSF17 (tumor necrosis factor receptor superfamily member 17) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). TNFRSF17 is also known in the art as B Cell Maturation Antigen (BCMA). The term encompasses "full-length" unprocessed TNFRSF17, as well as any form of TNFRSF17 resulting from processing in a cell. The term also encompasses naturally occurring variants of TNFRSF17, such as splice variants or allelic variants. The nucleic acid sequence of exemplary human TNFRSF17 is set forth in SEQ ID NO 19 (NCBI reference: NM-001192.3). The amino acid sequence of an exemplary protein encoded by human TNFRSF17 is shown in SEQ ID NO:20 (UNIPROT) ^TM Accession number Q02223-1).

The term "IGJ" as used herein, unless otherwise indicated, refers to any native IGJ (immunoglobulin J chain) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term includes "full-length" unprocessed IGJ, as well as any form of IGJ produced by processing in a cell. The term also encompasses naturally occurring variants of IGJ, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human IGJ is set forth in SEQ ID NO:21 (NCBI reference sequence: NM-144646.4). The amino acid sequence of an exemplary protein encoded by human IGJ is shown in SEQ ID NO 22 (UNIPROT) ^TM Accession number P01591-1).

The term "IGLL5" as used herein, unless otherwise indicated, refers to any native IGLL5 (immunoglobulin lambda-like polypeptide 5) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). IGLL5 is also known in the art as IGL, IGLV, and VL MAR. The term encompasses "full-length" unprocessed IGLL5, as well as any form of IGLL5 that results from processing in a cell. The term also encompasses naturally occurring variants of IGLL5, such as splice variants or allelic variants. The nucleic acid sequence of exemplary human IGLL5 is set forth in SEQ ID NO:23 (NCBI reference sequence: NM-001178126.2). The amino acid sequence of an exemplary protein encoded by human IGLL5 is shown in SEQ ID NO 24 (UNIPROT) ^TM Accession number B9A 064-1).

The term "RBPJ" as used herein, unless otherwise indicated, refers to any native RBPJ (recombinant signal binding protein of immunoglobulin kappa J region) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). RBPJ is also known in the art as CBF1 and hairless recombinant binding protein inhibitor. The term encompasses "full-length" unprocessed PRBJs, as well as any form of RBPJ resulting from processing in a cell. The term also encompasses naturally occurring variants of RBPJ, such as splice variants or allelic variants. Core of an exemplary human RBPJThe sequence is set forth in SEQ ID NO. 25 (NCBI reference sequence: NM-005349.3). The amino acid sequence of an exemplary protein encoded by human RBPJ is shown in SEQ ID NO 26 (UNIPROT) ^TM Accession number Q06330-1).

The term "MZB1" as used herein, unless otherwise indicated, refers to any native MZB1 (marginal zone B and B1 cell-specific proteins) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). MZB1 is also known in the art as MEDA-7, PACAP, and pERp1. The term encompasses "full-length" unprocessed MZB1, as well as any form of MZB1 that results from processing in the cell. The term also encompasses naturally occurring variants of MZB1, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human MZB1 is set forth in SEQ ID NO:27 (NCBI reference: NM-016459.4). The amino acid sequence of an exemplary protein encoded by human MZB1 is shown in SEQ ID NO 28 (UNIPROT) ^TM Accession number Q8WU 39-1).

The term "CCL2" as used herein, unless otherwise indicated, refers to any native CCL2 (chemokine (C-C motif) ligand 2) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). CCL2 is also known in the art as monocyte chemotactic protein 1 (MCP 1) and small inducible cytokine A2. The term encompasses "full-length" unprocessed CCL2, as well as any form of CCL2 that results from processing in a cell. The term also encompasses naturally occurring variants of CCL2, such as splice variants or allelic variants. The nucleic acid sequence of exemplary human CCL2 is set forth in SEQ ID NO:29 (NCBI reference sequence: NM-002982.4). The amino acid sequence of an exemplary protein encoded by human CCL2 is shown in SEQ ID NO 30 (UNIPROT) ^TM Accession number P13500-1).

The term "CCL3" as used herein, unless otherwise indicated, refers to any native CCL3 (chemokine (C-C motif) ligand 3) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). CCL 3 is also known in the art as macrophage inflammatory protein 1-alpha (MIP-1-alpha). The term encompasses "full-length" unprocessed CCL3, as well as any form of CCL3 that results from processing in a cell. The term also encompasses naturally occurring variants of CCL3, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human CCL3 is set forth in SEQ ID NO:31 (NCBI reference: NM-002983.3). The amino acid sequence of an exemplary protein encoded by human CCL3 is shown in SEQ ID NO 32 (UNIPROT) ^TM Accession number P10147-1).

The term "CCL4" as used herein, unless otherwise indicated, refers to any native CCL4 (chemokine (C-C motif) ligand 4) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). CCL4 is also known in the art as macrophage inflammatory protein 1-beta (MIP-1-beta). The term encompasses "full-length" unprocessed CCL4, as well as any form of CCL4 that results from processing in a cell. The term also encompasses naturally occurring variants of CCL4, such as splice variants or allelic variants. The nucleic acid sequence of exemplary human CCL4 is set forth in SEQ ID NO:33 (NCBI reference sequence: NM-002984.4). The amino acid sequence of an exemplary protein encoded by human CCL4 is shown in SEQ ID NO 34 (UNIPROT) ^TM Accession number P13236-1).

The term "CCL5" as used herein, unless otherwise indicated, refers to any native CCL5 (chemokine (C-C motif) ligand 5) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). CCL5 is also known in the art as modulating activating normal T cell expression secretion factors (RANTES), SCYA5, SIS- δ, SISd, TCP228, and eoCP. The term encompasses "full-length" unprocessed CCL5, as well as any form of CCL5 that results from processing in a cell. The term also encompasses naturally occurring variants of CCL5, such as splice variants or allelic variants. The nucleic acid sequence of exemplary human CCL5 is set forth in SEQ ID NO:35 (European nucleotide archive accession number AF 043341.1). The amino acid sequence of an exemplary protein encoded by human CCL5 is shown in SEQ ID NO 36 (UNIPROT) ^TM Accession number P13501-1).

The term "CCL8" as used herein, unless otherwise indicated, refers to any native CCL8 (chemokine (C-C motif) ligand 8) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). CCL8 is also known in the art as monocyte chemotactic protein 2 (MCP 2). The term encompasses "full-length" unprocessed CCL8, as well as any form of CCL8 that results from processing in a cell. The term also encompasses naturally occurring variants of CCL8, such as splice variants or allelic variants. The nucleic acid sequence of exemplary human CCL8 is set forth in SEQ ID NO:37 (NCBI reference sequence: NM-005623.3). The amino acid sequence of an exemplary protein encoded by human CCL8 is shown in SEQ ID NO 38 (UNIPROT) ^TM Accession number P80075-1).

The term "CCL18" as used herein, unless otherwise indicated, refers to any native CCL18 (chemokine (C-C motif) ligand 18) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). CCL18 is also known in the art as lung activation-regulated chemokine (PARC), dendritic Cell (DC) chemokine 1 (DC-CK 1), surrogate macrophage activation-related CC chemokine-1 (AMAC-1), and macrophage inflammatory protein-4 (MIP-4). The term encompasses "full-length" unprocessed CCL18, as well as any form of CCL18 that results from processing in a cell. The term also encompasses naturally occurring variants of CCL18, such as splice variants or allelic variants. The nucleic acid sequence of exemplary human CCL18 is set forth in SEQ ID NO:39 (NCBI reference sequence: NM-002988.4). The amino acid sequence of an exemplary protein encoded by human CCL18 is shown in SEQ ID NO 40 (UNIPROT) ^TM Accession number P55774-1).

The term "CCL19" as used herein, unless otherwise indicated, refers to any native CCL19 (chemokine (C-C motif) ligand 19) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). CCL19 is also in the art Known as EBI1 ligand chemokine (ELC) and macrophage inflammatory protein-3-beta (MIP-3-beta). The term encompasses "full-length" unprocessed CCL19, as well as any form of CCL19 that results from processing in a cell. The term also encompasses naturally occurring variants of CCL19, such as splice variants or allelic variants. The nucleic acid sequence of exemplary human CCL19 is set forth in SEQ ID NO:41 (NCBI reference sequence: NM-006274.3). The amino acid sequence of an exemplary protein encoded by human CCL19 is shown in SEQ ID NO 42 (UNIPROT) ^TM Accession number Q99731-1).

The term "CCL21" as used herein, unless otherwise indicated, refers to any native CCL21 (chemokine (C-C motif) ligand 21) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). CCL21 is also known in the art as 6Ckine, exodus-2 and secondary lymphoid tissue chemokine (SLC). The term encompasses "full-length" unprocessed CCL21, as well as any form of CCL21 that results from processing in a cell. The term also encompasses naturally occurring variants of CCL21, such as splice variants or allelic variants. The nucleic acid sequence of exemplary human CCL21 is set forth in SEQ ID NO 43 (NCBI reference: NM-002989.4). The amino acid sequence of an exemplary protein encoded by human CCL21 is shown in SEQ ID NO 44 (UNIPROT) ^TM Accession number O00585-1).

The term "CXCL9" as used herein, unless otherwise indicated, refers to any native CXCL9 (chemokine (C-X-C motif) ligand 9) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). CXCL9 is also known in the art as gamma interferon-induced Monokine (MIG). The term encompasses "full-length" unprocessed CXCL9, as well as any form of CXCL9 that results from processing in a cell. The term also encompasses naturally occurring variants of CXCL9, such as splice variants or allelic variants. The nucleic acid sequence of exemplary human CXCL9 is set forth in SEQ ID NO:45 (NCBI reference: NM-002416.3). The amino acid sequence of an exemplary protein encoded by human CXCL9 is shown in SEQ ID NO 46 (UNIPROT) ^TM Logging inNumber Q07325-1).

The term "CXCL10" as used herein, unless otherwise indicated, refers to any native CXCL10 (C-X-C motif chemokine ligand 10) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). CXCL10 is also known in the art as interferon gamma inducible protein 10 (IP-10) or small inducible cytokine B10. The term encompasses "full-length" unprocessed CXCL10, as well as any form of CXCL10 that results from processing in a cell. The term also encompasses naturally occurring variants of CXCL10, such as splice variants or allelic variants. The nucleic acid sequence of exemplary human CXCL10 is set forth in SEQ ID NO:47 (NCBI reference: NM-001565.4). The amino acid sequence of an exemplary protein encoded by human CXCL10 is shown in SEQ ID NO 48 (UNIPROT) ^TM Accession number P02778-1).

The term "CXCL11" as used herein, unless otherwise specified, refers to any native CXCL11 (C-X-C motif chemokine ligand 11) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). CXCL11 is also known in the art as interferon-inducible T cell alpha chemoattractant (I-TAC) and interferon-gamma inducible protein 9 (IP-9). The term encompasses "full-length" unprocessed CXCL11, as well as any form of CXCL11 that results from processing in a cell. The term also encompasses naturally occurring variants of CXCL11, such as splice variants or allelic variants. The nucleic acid sequence of exemplary human CXCL11 is set forth in SEQ ID NO:49 (NCBI reference sequence: NM-005409.5). The amino acid sequence of an exemplary protein encoded by human CXCL11 is shown in SEQ ID NO:50 (UNIPROT) ^TM Accession number O14625-1).

The term "CXCL13" as used herein, unless otherwise specified, refers to any native CXCL13 (C-X-C motif chemokine ligand 13) from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). CXCL13 is also known in the art as B Lymphocyte Chemoattractant (BLC) and B cell attraction Chemokine 1 (BCA-1). The term encompasses "full-length" unprocessed CXCL13, as well as any form of CXCL13 that results from processing in a cell. The term also encompasses naturally occurring variants of CXCL13, such as splice variants or allelic variants. The nucleic acid sequence of exemplary human CXCL13 is set forth in SEQ ID NO:51 (NCBI reference sequence: NM-006419.2). The amino acid sequence of an exemplary protein encoded by human CXCL13 is shown in SEQ ID NO 52 (UNIPROT) ^TM Accession number O43927-1).

Unless otherwise indicated, the term "CD8A" as used herein refers to a cluster of differentiated 8A genes, including any native CD8A from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length" unprocessed CD8A, as well as any form of CD8A resulting from processing in a cell. The term also encompasses naturally occurring variants of CD8A, such as splice variants or allelic variants. Exemplary nucleic acid sequence of human CD8A is set forth in SEQ ID NO 53 (GENBANK) ^TM Accession number M12828.1). The amino acid sequence of an exemplary protein encoded by human CD8A is shown in SEQ ID NO 54 (UNIPROT) ^TM Accession number P01732-1).

Unless otherwise indicated, the term "EOMES" as used herein refers to an apoprotein gene, including any native EOMES from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). EOMES is also known in the art as T-box brain protein 2 (Tbr 2). The term encompasses "full-length" unprocessed EOMES, as well as any form of EOMES that results from processing in a cell. The term also encompasses naturally occurring variants of EOMES, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human EOMES is set forth in SEQ ID NO:55 (NCBI reference: NM-005442.4). The amino acid sequence of an exemplary protein encoded by human EOMES is shown in SEQ ID NO 56 (UNIPROT) ^TM Accession number O95936-1).

The term "GZMA" as used herein, unless otherwise indicated, refers to the granzyme a gene, including from any ridgeAny native GZMA of vertebrate origin, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length" unprocessed GZMA, as well as any form of GZMA that results from processing in a cell. The term also encompasses naturally occurring variants of GZMA, such as splice variants or allelic variants. Exemplary nucleic acid sequences of human GZMA are set forth in SEQ ID NO:57 (GENBANK) ^TM Accession number BC 015739). The amino acid sequence of an exemplary protein encoded by human GZMA is shown in SEQ ID NO:58 (UNIPROT) ^TM Accession number P12544-1).

Unless otherwise indicated, the term "TBX21" as used herein refers to the T-box transcription factor TBX21 gene, including any native TBX21 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). TBX21 is also known in the art as T-PET, T-beta, TBLYM and T-box21. The term encompasses "full-length" unprocessed TBX21, as well as any form of TBX21 that results from processing in a cell. The term also encompasses naturally occurring variants of TBX21, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human TBX21 is set forth in SEQ ID NO:59 (NCBI reference sequence: NM-013351.2). The amino acid sequence of an exemplary protein encoded by human TBX21 is shown in SEQ ID NO:60 (UNIPROT) ^TM Accession number Q9UL 17-1).

Unless otherwise indicated, the term "IFNG" as used herein refers to the interferon gamma gene, including any native IFNG from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). IFNG is also known in the art as type II interferon. The term encompasses "full-length" unprocessed IFNG, as well as any form of IFNG that results from processing in a cell. The term also encompasses naturally occurring variants of IFNG, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human IFNG is set forth in SEQ ID NO:61 (NCBI reference sequence: NM-000619.3). The amino acid sequence of an exemplary protein encoded by human IFNG is shown in SEQ ID NO: 62(UNIPROT ^TM Accession number P01579-1).

The term "GZMB," as used herein, unless otherwise indicated, refers to the granzyme B gene, including any native GZMB from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length" unprocessed GZMB, as well as any form of GZMB that results from processing in a cell. The term also encompasses naturally occurring variants of GZMB, such as splice variants or allelic variants. Exemplary nucleic acid sequences of human GZMB are set forth in SEQ ID NO:63 (GENBANK) ^TM Accession number J03072). The amino acid sequence of an exemplary protein encoded by human GZMB is shown in SEQ ID NO 64 (UNIPROT) ^TM Accession number P10144-1).

The term "DERL3" as used herein, unless otherwise indicated, refers to the derlin-3 gene, including any native DERL3 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length" unprocessed DERL3, as well as any form of DERL3 that results from processing in a cell. The term also encompasses naturally occurring variants of DERL3, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human DERL3 is set forth in SEQ ID NO:65 (European nucleotide archive accession No. AK 125830.1). The amino acid sequence of an exemplary protein encoded by human DERL3 is shown in SEQ ID NO 66 (UNIPROT) ^TM Accession number Q96Q 80-1).

The term "JSRP1" as used herein, unless otherwise specified, refers to the connecting sarcoplasmic reticulin 1 gene, including any native JSRP1 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length" unprocessed JSRP1, as well as any form of JSRP1 that results from processing in a cell. The term also encompasses naturally occurring variants of JSRP1, such as splice variants or allelic variants. An exemplary nucleic acid sequence of human JSRP1 is set forth in SEQ ID NO 67 (European nucleotides)Archive accession number BC 021201.2). The amino acid sequence of an exemplary protein encoded by human JSRP1 is shown in SEQ ID NO 68 (UNIPROT) ^TM Accession number Q96MG 2-1).

Unless otherwise indicated, the term "IGHG2" as used herein refers to an immunoglobulin heavy chain constant γ 2 gene, including any native IGHG2 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length" unprocessed IGHG2, as well as any form of IGHG2 that results from processing in a cell. The term also encompasses naturally occurring variants of IGHG2, such as splice variants or allelic variants. The nucleic acid sequence of exemplary human IGHG2 is set forth in SEQ ID NO:69 (European nucleotide archive accession number AL 928742.3). The amino acid sequence of an exemplary protein encoded by human IGHG2 is shown in SEQ ID NO 70 (UNIPROT) ^TM Accession number P01859-1).

The term "IGHGP" as used herein, unless otherwise indicated, refers to an immunoglobulin heavy chain constant gamma P gene, including any native IGHGP from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length" unprocessed IGHGP, as well as any form of IGHGP that results from processing in a cell. The term also encompasses naturally occurring variants of IGHGP, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human IGHGP is set forth in SEQ ID NO:71 (NCBI reference sequence No. NG _ 001019.6).

The term "IGLV3-1" as used herein, unless otherwise indicated, refers to an immunoglobulin λ variable 3-1 gene, including any native IGLV3-1 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length" unprocessed IGLV3-1, as well as any form of IGLV3-1 that results from processing in the cell. The term also encompasses naturally occurring variants of IGLV3-1, such as splice variants or allelic variants. Exemplary human IGLV3-1 nucleus The sequence is set forth in SEQ ID NO:72 (European nucleotide archive accession No. AC 2458028.2). The amino acid sequence of an exemplary protein encoded by human IGLV3-1 is shown in SEQ ID NO 73 (UNIPROT) ^TM Accession number P01715-1).

The term "IGLV6-57" as used herein, unless otherwise indicated, refers to an immunoglobulin λ variable 6-57 gene, including any native IGLV6-57 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length" unprocessed IGLV6-57, as well as any form of IGLV6-57 that results from processing in the cell. The term also encompasses naturally occurring variants of IGLV6-57, such as splice variants or allelic variants. The nucleic acid sequence of exemplary human IGLV6-57 is set forth in SEQ ID NO:74 (European nucleotide archive accession number AC 245760.1). The amino acid sequence of an exemplary protein encoded by human IGLV6-57 is shown in SEQ ID NO 75 (UNIPROT) ^TM Accession number P01721-1).

Unless otherwise indicated, the term "IGHA2" as used herein refers to an immunoglobulin heavy chain constant α 2 gene, including any native IGHG2 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length" unprocessed IGHA2, as well as any form of IGHA2 that results from processing in a cell. The term also encompasses naturally occurring variants of IGHA2, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human IGHA2 is set forth in SEQ ID NO:76 (European nucleotide archive accession number AL 928742.3). The amino acid sequence of an exemplary protein encoded by human IGHA2 is shown in SEQ ID NO 77 (UNIPROT) ^TM Accession number P01877-1).

Unless otherwise indicated, the term "IGKV4-1" as used herein refers to immunoglobulin kappa variable 4-1 genes, including any native IGKV4-1 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses the absence of "full lengthIndustrial IGKV4-1, and any form of IGKV4-1 that results from processing in cells. The term also encompasses naturally occurring variants of IGKV4-1, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human IGKV4-1 is set forth in SEQ ID NO:78 (European nucleotide archive accession number X02990.1). The amino acid sequence of an exemplary protein encoded by human IGKV4-1 is shown in SEQ ID NO:79 (UNIPROT) ^TM Accession number P06312-1).

The term "IGKV1-12" as used herein, unless otherwise indicated, refers to immunoglobulin kappa variable 1-12 genes, including any native IGKV1-12 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length" unprocessed IGKV1-12, as well as any form of IGKV1-12 that results from processing in a cell. The term also encompasses naturally occurring variants of IGKV1-12, such as splice variants or allelic variants. The nucleic acid sequence of an exemplary human IGKV1-12 is set forth in SEQ ID NO:80 (European nucleotide archive accession number AC 24579.2). The amino acid sequence of an exemplary protein encoded by human IGKV1-12 is shown in SEQ ID NO 81 (UNIPROT) ^TM Accession number A0A0C4DH 73-1).

The term "IGLC7" as used herein, unless otherwise indicated, refers to the immunoglobulin λ constant 7 gene, including any native IGLC7 from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term encompasses "full-length" unprocessed IGLC7, as well as any form of IGLC7 that results from processing in a cell. The term also encompasses naturally occurring variants of IGLC7, such as splice variants or allelic variants. The nucleic acid sequence of exemplary human IGLC7 is set forth in SEQ ID NO:82 (European nucleotide archive accession number AC 245973.2). The amino acid sequence of an exemplary protein encoded by human IGLC7 is shown in SEQ ID NO 83 (UNIPROT) ^TM Accession number A0M8Q 6-1).

The term "clonally expanded B cells" refers to B cells that have a common antigen specificity as assessed by, for example, sequence homology to the migg portion of the BCR. For example, clonally expanded B cells may share at least 85% sequence identity (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.9%, or 100% sequence identity) in their mIg portion heavy and/or light chains, e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.9%, or 100% sequence identity in the CDR1, CDR2, and/or CDR3 regions of their migg portion heavy and/or light chains.

The terms "cancer" and "cancerous" refer to or describe the physiological condition in mammals that is typically characterized by uncontrolled cell growth. Examples of cancer include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia or lymphoid malignancies. More specific examples of such cancers include, but are not limited to: lung cancer, including small cell lung cancer, NSCLC (e.g., non-squamous NSCLC and squamous NSCLC), adenocarcinoma of the lung, and squamous carcinoma of the lung; bladder cancer (e.g., urothelial Cancer (UC), muscle Invasive Bladder Cancer (MIBC), and BCG refractory non-muscle invasive bladder cancer (NMIBC)); kidney or renal cancer (e.g., renal Cell Carcinoma (RCC)); urinary tract cancer; breast cancer (e.g., HER2+ breast cancer and Triple Negative Breast Cancer (TNBC), which refers to estrogen receptor (ER-), progestin receptor (PR-) and HER2 (HER 2-) being negative); prostate cancer, such as castration-resistant prostate cancer (CRPC); peritoneal cancer; hepatocellular carcinoma; gastric cancer (gastric/gastric cancer), including gastrointestinal and gastrointestinal stromal cancer; pancreatic cancer; glioblastoma; cervical cancer; ovarian cancer; liver cancer; hepatoma; colon cancer; rectal cancer; carcinoma of large intestine; endometrial or uterine cancer; salivary gland cancer; prostate cancer; vulvar cancer; thyroid cancer; liver cancer; anal cancer; penile cancer; melanomas, including superficial invasive melanoma, malignant lentigo melanoma, peripheral macular malignant melanoma, and nodular melanoma; multiple myeloma and B-cell lymphomas (including low grade/follicular non-Hodgkin's lymphoma (NHL); small Lymphocytic (SL) NHL; moderate/follicular NHL; moderate diffuse NHL; hyperimmunoblastic NHL; high lymphocytic NHL; high small non-lytic NHL; giant lump NHL; mantle cell lymphoma; AIDS-related lymphoma; and Waldenstrom's Macroglobulinemia)); chronic Lymphocytic Leukemia (CLL); acute Lymphocytic Leukemia (ALL); acute Myeloid Leukemia (AML); hairy cell leukemia; chronic Myeloid Leukemia (CML); post-transplant lymphoproliferative disorder (PTLD); and myelodysplastic syndrome (MDS), as well as abnormal vascular proliferation associated with lentigo hamartoma, edema (such as diseases associated with brain tumors), meigs' syndrome, brain cancer, head and neck cancer, and related metastases.

The terms "cell proliferative disease" and "proliferative disease" refer to a condition associated with a degree of abnormal cell proliferation. In one embodiment, the cell proliferative disorder is cancer (e.g., lung cancer (e.g., NSCLC, including non-squamous NSCLC and squamous NSCLC), bladder cancer (e.g., UC), renal cancer (e.g., RCC), or breast cancer (e.g., TNBC)). In another embodiment, the cell proliferative disorder is a tumor.

A "chemotherapeutic agent" is a compound useful for treating cancer (e.g., lung cancer (e.g., NSCLC, including non-squamous NSCLC and squamous NSCLC), bladder cancer (e.g., UC), renal cancer (e.g., RCC), or breast cancer (e.g., TNBC)). Examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclophosphamide

Alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzotepa, carbaquinone, metodepa and uredepa; vinyl imines and methyl melamines, including tritylamine, triethylphosphoramide, triethylenethiophosphoramide, and trimethylolmelamine; polyacetylenyl (acetogenin, especially bucindocin and bucindolone); delta-9-tetrahydrocannabinol (dronabinol,

) (ii) a Beta-lapachone; lapachol; colchicine; betulinic acid; camptothecin (including the synthetic analogue topotecan)

CPT-11 (irinotecan,

) Acetyl camptothecin, scopoletin and 9-aminocamptothecin); bryostatins; cariostatin (callystatin); CC-1065 (including its aldorexin, kazelaixin, and bizelaixin synthetic analogs); podophyllotoxin; podophyllinic acid; teniposide; nostoc (especially nostoc 1 and nostoc 8); dolastatin; duocarmycins (including synthetic analogs, KW-2189 and CB1-TM 1); eleutherobin; (ii) coprinus atramentarius alkali; alcohol of coral tree; sponge chalone; nitrogen mustards such as chlorambucil, chlornaphazine, chlorophosphoramide, estramustine, ifosfamide, mechlorethamine oxide hydrochloride, melphalan, neonebixin, benzene mustard cholesterol, prednimustine, trofosfamide, uramustine; nitrosoureas such as carmustine, chlorourethrin, fotemustine, lomustine, nimustine and ranimustine; antibiotics, such as enediyne antibiotics (e.g., calicheamicin, particularly calicheamicin γ 1 'and calicheamicin ω 1' (see, e.g., nicolaou et al, angew. Chem Intl. Ed. Engl.,33, 183-186 (1994)); CDP323, oral α -4 integrin inhibitors, dalcomycin, including dalcomycin A, esperamicin, and neocarcinomycin chromophores and related tryptophane diyne antibiotic chromophores), acrinomycin, actinomycin, antromycin, azalomycin, azaserine, bleomycin, actinomycin C, karabixin, carminomycin, carvachin, tryptophacin, tryptophycin, actinomycin D, daunorubicin, ditorexin, 6-diazo-5-oxo-norleucine, doxorubicin, including doxorubicin

Morpholino-doxorubicin, cyano-morpholino-doxorubicin, 2-pyrroline-doxorubicin, doxorubicin hydrochloride liposome injection

LipidDoxorubicin TLC D-99

Pegylated liposomal doxorubicin

And doxorubicine), epirubicin, esorubicin, idarubicin, sisomicin, mitomycins such as mitomycin C, mycophenolic acid, nogomycin, olivomycin, pellomycin, pofimycin, puromycin, triiron doxorubicin, nodubicin, streptonigrin, streptozotocin, tubercidin, ubenimex, setastatin, zorubicin; antimetabolites, such as methotrexate, gemcitabine

Tegafur

Capecitabine

Epothilone and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, arabinoside, dideoxyuridine, doxifluridine, enocitabine, and floxuridine; androgens such as carpoterone, drotandrosterone propionate, epitioandrostanol, meindrotane, testolactone; anti-adrenalines, such as aminoglutethimide, mitotane, trostane; folic acid replenisher such as folinic acid; d, D-glucuronolactone acetate; an aldphosphoramide glycoside; (ii) aminolevulinic acid; eniluracil; amsacrine; amoxicillin; a bisantrene group; edatrexae; 1, ground Budd famine; colchicine; diazaquinone; (ii) nilotinib; (ii) hydroxypyrazole acetate; an epothilone; etoglut; gallium nitrate; a hydroxyurea; lentinan; lonidamine; maytansinoids such as maytansine and ansamitocins; mitoxantrone Hydrazone; mitoxantrone; mopidamole; diamine nitracridine; pentostatin; methionine; pirarubicin; losoxanthraquinone; 2-ethyl hydrazide; methylbenzyl hydrazine;

polysaccharide complexes (JHS Natural Products, eugene, OR); lezoxan; rhizomycin; a texaphyrin; a germanium spiroamine; alternarionic acid; a tri-imine quinone; 2,2' -trichlorotriethylamine; trichothecene toxins (particularly T-2 toxin, verrucosin A, bacillocin A and serpentine); uratan; vindesine

Dacarbazine; mannitol mustard; dibromomannitol; dibromodulcitol; pipobroman; geseiguxin; cytarabine ("Ara-C"); thiotepa triamine; the taxane class(s) is (are), such as paclitaxel (A)

Albumin engineered nanoparticle formulation of Bristol-Myers Squibb Oncology, princeton, N.J.), paclitaxel (ABRAXANE) ^TM ) And docetaxel (c), (d)

Rhome-Poulene Rorer, antony, france), chlorambucil; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs, such as cisplatin, platinum oxalate (e.g.,

) And carboplatin; catharanthus roseus for preventing tubulin polymerization to form microtubules, including vinblastine

Vincristine

Vindesine

And vinorelbine

Etoposide (VP-16); an ifosfamide; mitoxantrone; aldehyde hydrofolic acid; noscapine; edatrexae; daunomycin; aminopterin; iban phosphate; topoisomerase inhibitor RFS 2000; difluoromethyl ornithine (DMFO); retinoids such as retinoic acid, including bexarotene

Diphosphoric hydrochloric acids, such as chlorophosphate (e.g.,

or

) Etidrophosphates of formula (I)

NE-58095, zoledronic acid/zoledronic acid salt

Alendronate

Pamidronate salt

Tiluphosphate salt

Or risedronate

And troxacitabine (1, 3-dioxolane nucleoside cytosine analogues); antisense oligonucleotides, particularly those that inhibit the expression of genes in signaling pathways involved in abnormal cell proliferation, can decrease cell proliferation, e.g., PKC-alpha, raf, H-Ras and epidermal growth factor receptor (EGF-R) (examples)Such as erlotinib (TARCEVA) ^TM ) ); and VEGF-A; vaccines, e.g.

Vaccines and gene therapy vaccines, e.g.

A vaccine,

Vaccines and

a vaccine; a topoisomerase 1 inhibitor (for example,

) (ii) a The rmRH (for example,

) (ii) a BAY439006 (Sorafenib; bayer); SU-11248 (sunitinib,

pfizer); pirifoxin, COX-2 inhibitors (e.g., celecoxib or etoxib), proteasome inhibitors (e.g., PS 341); bortezomib

CCI-779; tipifarnib (R11577); sorafenib, ABT510; bcl-2 inhibitors such as sodium orbellison

Pixantrone; an EGFR inhibitor; tyrosine kinase inhibitors; serine-threonine kinase inhibitors such as rapamycin (sirolimus,

) (ii) a Farnesyl transferase inhibitors such as Lonafarnib (SCH 6636, sarasar) ^TM ) (ii) a And any pharmaceutically acceptable salt, acid or derivative thereof as described above;and combinations of two or more of the above, such as CHOP (abbreviation for cyclophosphamide, doxorubicin, vincristine and prednisolone combination therapy) and FOLFOX (oxaliplatin) ^TM ) Abbreviations for therapeutic regimens combining 5-FU and folinic acid), as well as any pharmaceutically acceptable salts, acids or derivatives thereof, described above; and combinations of two or more of the foregoing.

Chemotherapeutic agents as defined herein also include "anti-hormonal agents" or "endocrine therapeutic agents" for modulating, reducing, blocking or inhibiting the effects of hormones that can promote the growth of cancer (e.g., lung cancer (e.g., NSCLC, including non-squamous NSCLC and squamous NSCLC), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), or breast cancer (e.g., TNBC)). They may themselves be hormones, including but not limited to: antiestrogens and Selective Estrogen Receptor Modulators (SERMs), including, for example, tamoxifen (including

Tamoxifen), raloxifene, droloxifene, 4-hydroxyttamoxifen, troxifene, keoxifene, LY117018, onapristone and

(toremifene); aromatase inhibitors which inhibit aromatase, which regulates the production of estrogen in the adrenal glands, such as 4 (5) -imidazoles, aminoglutarimides,

Megestrol acetate,

Exemestane, formestane (formastanie), fadrozole,

Vorozole (vorozole),

Letrozole and

anastrozole; and antiandrogens such as flutamide, nilutamide, bicalutamide, leuprorelin and goserelin; and troxacitabine (1, 3-dioxolane nucleoside analog); antisense oligonucleotides, particularly those that inhibit gene expression in signaling pathways implicated by abnormal cell proliferation, such as PKC- α, raf and H-Ras; ribozymes, such as VEGF expression inhibitors (e.g.,

ribozymes) and inhibitors of HER2 expression; vaccines, such as gene therapy vaccines, e.g.

A vaccine,

Vaccines and

a vaccine;

rIL-2；

a topoisomerase 1 inhibitor;

rmRH; vinorelbine (vinorelbine) and esperamicins (esperamicins) (see U.S. Pat. No. 4,675,187); and pharmaceutically acceptable salts, acids or derivatives of any of the foregoing; and combinations of two or more of the foregoing.

The term "chimeric" antibody refers to an antibody in which a portion of the heavy and/or light chain is derived from a particular source or species, while the remainder of the heavy and/or light chain is derived from a different source or species.

"class" of antibody refers toThe type of constant domain or constant region that the heavy chain has. There are five major classes of antibodies: igA, igD, igE, igG and IgM, and some of these antibodies may be further divided into subclasses (isotypes), e.g., igG ₁ 、IgG ₂ 、IgG ₃ 、IgG ₄ 、IgA ₁ And IgA ₂ . The heavy chain constant domains corresponding to different classes of immunoglobulins are referred to as α, δ, ε, γ, and μ, respectively.

As used herein, the term "cytotoxic agent" refers to a substance that inhibits or prevents cellular function and/or causes cell death or destruction. Cytotoxic agents include, but are not limited to, radioisotopes (e.g., at) ²¹¹ 、I ¹³¹ 、I ¹²⁵ 、Y ⁹⁰ 、Re ¹⁸⁶ 、Re ¹⁸⁸ 、Sm ¹⁵³ 、Bi ²¹² 、P ³² 、Pb ²¹² And radioactive isotopes of Lu); chemotherapeutic agents or drugs (e.g., methotrexate, doxorubicin, vinca alkaloids (vincristine, vinblastine, etoposide), doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin, or other intercalating agents); a growth inhibitor; enzymes and fragments thereof such as nucleolytic enzymes; an antibiotic; toxins such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and/or variants thereof; and various anti-tumor or anti-cancer agents disclosed below.

The term "concurrently" as used herein refers to administration of two or more therapeutic agents, wherein at least some of the administrations overlap in time. Thus, concurrent administration includes a dosing regimen that continues administration of one or more additional agents after discontinuing administration of one or more agents.

As used herein, "delaying" the progression of a condition or disease means delaying, impeding, slowing, stabilizing, and/or delaying the progression of the disease or condition (e.g., cancer, such as lung cancer (e.g., NSCLC, including non-squamous NSCLC and squamous NSCLC), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), or breast cancer (e.g., TNBC)). Such delays may be of varying lengths of time, depending on the medical history and/or the subject to be treated. It will be apparent to those skilled in the art that a sufficient or significant delay may actually encompass prevention, as the subject will not suffer from the disease.

The terms "determining", "detecting" and grammatical variations thereof include any means of determining or detecting, including direct and indirect determination or detection.

By "disorder" or "disease" is meant any condition that may benefit from treatment, including, but not limited to, chronic and acute disorders or diseases, including those pathological conditions that predispose a mammal to the disorder (e.g., cancer, such as lung cancer (e.g., NSCLC, including non-squamous NSCLC and squamous NSCLC), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), or breast cancer (e.g., TNBC)).

The term "diagnosis" as used herein refers to the determination or classification of a molecular or pathological condition, disease or disorder, such as cancer, e.g., lung cancer (e.g., NSCLC, including non-squamous NSCLC and squamous NSCLC), bladder cancer (e.g., UC), renal cancer (e.g., RCC), or breast cancer (e.g., TNBC). For example, "diagnosis" may refer to the identification of a particular type of cancer. "diagnosis" may also refer to the classification of a particular subtype of cancer, for example, by histopathological criteria or molecular features (e.g., a subtype characterized by expression of one or a combination of biomarkers (e.g., a particular gene or protein encoded by such gene)).

"Effector function" refers to those biological activities that can be attributed to the Fc region of an antibody that vary with the isotype of the antibody. Examples of antibody effector functions include: c1q binding and Complement Dependent Cytotoxicity (CDC); fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; down-regulation of cell surface receptors (e.g., PD-L1); and B cell activation.

An "effective amount" of a compound, e.g., a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., an atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) or a composition thereof (e.g., a pharmaceutical composition), is the minimum amount to achieve a desired therapeutic or prophylactic effect, such as a measurable increase in Overall Survival (OS), progression-free survival (PFS), or overall response (e.g., a determined optimal overall response (BCOR)) of a particular disease or condition (e.g., cancer, e.g., lung cancer (e.g., NSCLC, including non-squamous NSCLC and squamous NSCLC), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), or breast cancer (e.g., TNBC)). An effective amount herein may vary depending on factors such as the disease state, age, sex, and weight of the individual, and the ability of the antibody to elicit a desired response in the subject. An effective amount is also one in which any toxic or detrimental effects of the treatment are outweighed by the therapeutically beneficial effects. For prophylactic use, beneficial or desired results include, for example, elimination or reduction of risk, lessening of severity, or delaying onset of the disease, which includes biochemical, histological, and/or behavioral symptoms of the disease, complications thereof, and intermediate pathological phenotypes that arise during the course of disease development. The effective amount may be administered one or more times. For the purposes of this disclosure, an effective amount of a drug, compound or pharmaceutical composition is an amount sufficient for direct or indirect prophylaxis or treatment. As understood in the clinical setting, an effective amount of a drug, compound or pharmaceutical composition can optionally be achieved in combination with another drug, compound or pharmaceutical composition. Thus, an "effective amount" may be considered in the context of administering one or more therapeutic agents, and administration of an effective amount of a single agent may be considered if the desired result can be achieved or achieved in combination with one or more other agents. For example, an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) as a cancer treatment can reduce the number of cancer cells; reducing primary tumor size; inhibit (i.e., slow to some extent and preferably stop) cancer cell infiltration into peripheral organs; inhibit (i.e., slow and preferably stop to some extent) tumor metastasis; inhibit tumor growth to some extent; and/or relieve to some extent one or more symptoms associated with the condition. To the extent that the drug prevents growth and/or kills existing cancer cells, it can inhibit cell growth and/or be cytotoxic. For cancer therapy, in vivo efficacy can be measured, for example, by assessing survival time, time to disease progression (TTP), response Rate (RR), duration of response, and/or quality of life.

The term "Fc region" is used herein to define the C-terminal region of an immunoglobulin heavy chain, which comprises at least a portion of a constant region. The term includes native sequence Fc regions and variant Fc regions. In one embodiment, the human IgG heavy chain Fc region extends from Cys226 or from Pro230 to the carboxy terminus of the heavy chain. However, the C-terminal lysine (Lys 447) of the Fc region may or may not be present. Unless otherwise specified herein, the numbering of amino acid residues in the Fc region or constant region is according to the EU numbering system, also known as the EU index, as described in Kabat et al, sequences of Proteins of Immunological Interest, 5 th edition, public Health Service, national Institutes of Health, bethesda, MD, 1991.

"framework" or "FR" refers to variable domain residues other than hypervariable region (HVR) residues. The FRs of a variable domain typically consist of the following four FR domains: FR1, FR2, FR3 and FR4. Thus, HVR and FR sequences typically occur in the VH (or VL) as follows: FR1-H1 (L1) -FR2-H2 (L2) -FR3-H3 (L3) -FR4.

The terms "full-length antibody," "intact antibody," and "whole antibody" are used interchangeably herein to refer to an antibody having a structure substantially similar to a native antibody structure or having a heavy chain containing an Fc region as defined herein.

A "human antibody" is an antibody having an amino acid sequence corresponding to that of an antibody produced by a human or human cell, or derived from an antibody of non-human origin using a repertoire of human antibodies or other human antibody coding sequences. This definition of human antibody specifically excludes humanized antibodies comprising non-human antigen binding residues. Human antibodies, including phage display libraries, can be generated using a variety of techniques known in the art. Hoogenboom and Winter, j.mol.biol.,227 (1991); marks et al, j.mol.biol.,222 (1991). Also useful for the preparation of human Monoclonal Antibodies are methods such as Cole et al, monoclonal Antibodies and Cancer Therapy, alan R.Liss, p.77 (1985); boerner et al, J.Immunol.,147 (1): 86-95 (1991). See also van Dijk et al, curr, opin, pharmacol, 5. Human antibodies can be made by administering an antigen to a transgenic animal that has been modified to produce such antibodies in response to an antigen challenge but has failed at its endogenous locus, e.g., to immunize a XENOMOUSE (see, e.g., U.S. Pat. Nos. 6,075,181 and 6,150,584 for Xenomose ^TM A technique). See also, e.g., li et al, proc.natl.acad.sci.usa,103, 3557-3562 (2006) for human antibodies produced by human B cell hybridoma technology.

A "humanized" antibody is a chimeric antibody comprising amino acid residues from non-human HVRs and amino acid residues from human FRs. In certain embodiments, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the HVRs (e.g., CDRs) correspond to those of a non-human antibody, and all or substantially all of the FRs correspond to those of a human antibody. The humanized antibody optionally may comprise at least a portion of an antibody constant region derived from a human antibody. An antibody that is a "humanized form," e.g., a non-human antibody, refers to an antibody that has been humanized.

The term "hypervariable region" or "HVR" as used herein refers to each of the regions in the sequence that are hypervariable ("complementarity determining regions" or "CDRs") and/or form structurally defined loops ("hypervariable loops") and/or antibody variable domains containing antigen-contacting residues ("antigen-contacting points"). Typically, an antibody comprises six HVRs: three in VH (H1, H2, H3) and three in VL (L1, L2, L3). Exemplary HVRs herein include:

(a) Highly variable loops occurring at the following amino acid residues: 26 to 32 (L1), 50 to 52 (L2), 91 to 96 (L3), 26 to 32 (H1), 53 to 55 (H2) and 96 to 101 (H3) (Chothia and Lesk, J.mol.biol.196:901-917 (1987));

(b) CDRs occurring at amino acid residues 24-34 (L1), 50-56 (L2), 89-97 (L3), 31-35b (H1), 50-65 (H2) and 95-102 (H3) (Kabat et al, sequences of Proteins of Immunological Interest, 5 th edition, public Health Service, national Institutes of Health, bethesda, MD (1991));

(c) Antigen contacts occurring at amino acid residues 27c-36 (L1), 46-55 (L2), 89-96 (L3), 30-35b (H1), 47-58 (H2), and 93-101 (H3) (MacCallum et al, J.mol.biol.262:732-745 (1996)); and

(d) A combination of (a), (b), and/or (c) that comprises HVR amino acid residues 46-56 (L2), 47-56 (L2), 48-56 (L2), 49-56 (L2), 26-35 (H1), 26-35b (H1), 49-65 (H2), 93-102 (H3), and 94-102 (H3).

Unless otherwise indicated, HVR residues and other residues (e.g., FR residues) in the variable domains are numbered herein according to Kabat et al, supra.

An "isolated" antibody is one that has been separated from components of its natural environment. In some embodiments, the antibody is purified to greater than 95% or 99% purity as determined, for example, by electrophoresis (e.g., SDS-PAGE, isoelectric focusing (IEF), capillary electrophoresis), or chromatography (e.g., ion exchange or reverse phase HPLC). For a review of methods for assessing antibody purity, see, e.g., flatman et al, j.chromager.b 848 (2007).

An "isolated" nucleic acid is a nucleic acid molecule that has been separated from components of its natural environment. An isolated nucleic acid includes a nucleic acid molecule that is contained in a cell that normally contains the nucleic acid molecule, but which is present extrachromosomally or at a chromosomal location that is different from its natural chromosomal location.

As used herein, the word "label" refers to a detectable compound or composition. Labels are typically conjugated or fused, directly or indirectly, to an agent, such as a polynucleotide probe or antibody, and facilitate detection of the conjugated or fused agent. The label may be detectable by itself (e.g., radioisotope labels or fluorescent labels) or, in the case of an enzymatic label, may catalyze chemical alteration of a substrate compound or composition to produce a detectable product.

In general, the terms "level of expression" or "expression level" are used interchangeably and generally refer to the amount of a biomarker in a biological sample. "expression" generally refers to the process by which information (e.g., gene-encoded and/or epigenetic) is converted into structures present in and operating in a cell. Thus, as used herein, "expression" may refer to transcription into a polynucleotide, translation into a polypeptide, or even polynucleotide and/or polypeptide modifications (e.g., post-translational modifications of a polypeptide). Transcribed polynucleotides, translated polypeptides, or fragments of polynucleotide and/or polypeptide modifications (e.g., post-translational modifications of polypeptides) should also be considered as expressed, whether they are derived from transcripts generated by alternatively spliced or degraded transcripts, or from post-translational processing (e.g., by proteolysis) of polypeptides. "expressed genes" include those that are transcribed into a polynucleotide, such as an mRNA, and then translated into a polypeptide, and also those that are transcribed into RNA but not translated into a polypeptide (e.g., transfer RNA and ribosomal RNA). Expression levels can be measured by methods known to those skilled in the art and also disclosed herein, including, for example, RT-qPCR and RNA-seq. The assessed expression level can be used to determine a response to the treatment.

The term "immune score expression level" as used herein refers to a numerical value that reflects the expression level of a single gene of interest (e.g., normalized expression level) or the collective expression level of multiple genes of interest (e.g., at least two, at least three, at least four, at least five, or more genes of interest) that are associated with an immune response. The immune score expression level of more than one gene of interest can be determined by collective methods known to those skilled in the art and also disclosed herein, including, for example, by calculating the median or mean of the expression levels of all genes of interest. Prior to pooling, the expression level of each gene of interest can be normalized using statistical methods known to those skilled in the art and disclosed herein, including, for example, normalization to the expression level of one or more housekeeping genes, or to the total library size, or to the median or average expression level value of all measured genes. In some cases, the normalized expression level of each gene of interest may be normalized by calculating a Z-score for the normalized expression level of each gene of interest prior to the plurality of sets of genes of interest. In some cases, each gene of interest may have a weight score assigned, and the immune score expression level of a plurality of genes of interest may be calculated by combining the weight scores to determine a mean of the weighted expression levels of all genes of interest. For example, an immune score expression level can refer to a value reflecting the normalized expression level of a single gene selected from any one of tables 1 to 17 (e.g., CD79A, CD19, BANK1, jchanin, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, MZB1, CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, CXCL13, DERL3, JSRP1, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, and IGLC 7). Alternatively, the immune score expression level may, for example, refer to an expression level that reflects a mean (e.g., normalized) expression level of at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, or a normalized expression level (e.g., normalized expression level of at least 18, such as a normalized expression level or a normalized expression level of at least 18, such as a normalized expression level of a set of expression levels (e.g., a normalized expression level of at least 18, at least 19, or a normalized expression level of at least two, such as CD79A, CD19, BANK1, jchan 1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, rbj, mzj, MZB1, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, IGKV1, IGLC 11, IGLC 13, IGLC7, or a combination thereof). In some cases, an immune score expression level can, for example, refer to a level that reflects the expression of a gene listed in any one of tables 1 to 17 (e.g., CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, MZB1, CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, CXCL13, DERL3, JSRP1, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, and IGLC7, or combinations thereof) at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or more of the collective Z-score (e.g., the normalized Z-score or the normalized Z-score of the expression levels.

As used herein, the term "reference immune score expression level" refers to an immune score expression level to which another immune score expression level (e.g., for one or more of the genes listed in any one of tables 1 to 17 (e.g., CD79A, CD19, BANK1, jchanin, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, MZB1, CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, CXCL13, DERL3, JSRP1, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, and IGLC 7)) is compared to make, for example, a diagnostic, predictive, prognostic, and/or therapeutic decision. For example, a reference immune score expression level can be derived from a reference sample, an expression level in a reference population (e.g., for a gene listed in any one of tables 1-17 (e.g., one or more of CD79A, CD19, BANK1, jchanin, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, MZB1, CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, CXCL13, DERL3, JSRP1, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, and IGLC 7)) and/or a pre-specified value (e.g., a cut-off value that previously determined satisfies the following conditions: the cutoff value is based on a significant difference above and/or below the cutoff value between the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist therapy and the responsiveness of the individual to treatment with the non-PD-L1 axis binding antagonist therapy, the significant (e.g., statistical) difference distinguishing a first subset of individuals in a reference population that have received treatment with a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attrituximab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) from a second subset of individuals in the same reference population that have received treatment with a non-PD-L1 axis binding antagonist therapy that does not contain a PD-L1 axis binding antagonist, wherein the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist therapy is significantly (e.g., statistically) improved relative to the responsiveness of the individual to treatment with the non-PD-L1 axis binding antagonist therapy relative to the treatment with the cutoff value, and/or the responsiveness of the individual to treatment with the non-PD-L1 axis binding antagonist therapy is significantly (e.g., statistically) improved relative to the responsiveness to the non-PD-L1 axis binding antagonist therapy relative to the treatment An individual's responsiveness to treatment with a PD-L1 axis binding antagonist is significantly (e.g., statistically) improved below a threshold value). One skilled in the art will recognize that the numerical value of the reference immune score expression level may vary depending on the indication (e.g., cancer (e.g., breast cancer, lung cancer (e.g., NSCLC, including non-squamous NSCLC and squamous NSCLC), renal cancer, or bladder cancer), the method used to detect the expression level (e.g., RNA-seq or RT-qPCR), the statistical method used to generate the immune score, and/or the particular combination of genes examined.

By "increased expression", "increased expression level" or "increased level" is meant an increased level of expression of a gene or combination of genes (e.g., for one or more of the genes listed in any one of tables 1 to 17 (e.g., CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, MZB1, CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, CXCL13, DERL3, JSRP1, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, and IGLC 7)) in a subject relative to a control, such as an uninvolved disease or condition (e.g., a cancer, e.g., lung cancer (e.g., including non-NSCLC and NSCLC), bladder cancer (e.g., squamous cell carcinoma (e.g., renal carcinoma), e.g., a squamous cell carcinoma (e.g., a reference, e.g., a reference example, a squamous cell cancer) or a reference level, such as an immune or a reference example, a squamous cell, a breast cancer, a reference example, a.

By "reduced expression", "reduced expression level" or "reduced level" is meant a reduced level of expression of a gene or combination of genes (e.g., for one or more of the genes listed in any one of tables 1 to 17 (e.g., CD79A, CD19, BANK1, jchan, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, MZB1, CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, CXCL13, DERL3, JSRP1, IGHG2, IGHGP, igcl 3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, and IGLC 7)) in a subject relative to a control (such as an uninvolved disease or condition (e.g., a cancer, e.g., lung cancer (e.g., including non-NSCLC and NSCLC), bladder cancer (e.g., squamous cell carcinoma (e.g., TNBC), e.g., squamous cell carcinoma (e.g., a reference example, a reference level or a reference level of an immune or a reference example, such as a squamous cell cancer) or a reference example. In some embodiments, reduced expression is little or no expression.

As used herein, "reference gene" refers to a gene or set of genes (e.g., one, two, three, four, five, or six or more genes) for comparison purposes, such as a housekeeping gene. By "housekeeping gene" is meant herein a gene or set of genes (e.g., one, two, three, four, five, or six or more genes) that encodes a protein whose activity is critical for maintaining cell function and which is typically similarly present in all cell types.

The term "monoclonal antibody" as used herein refers to an antibody obtained from a substantially homogeneous population of antibodies, i.e., individual antibodies comprising the population share identity and/or bind the same epitope, except for possible variant antibodies (e.g., containing naturally occurring mutations or produced during the manufacture of a monoclonal antibody preparation, such variants typically being present in minor amounts). In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody in a monoclonal antibody preparation is directed against a single determinant on the antigen. Thus, the modifier "monoclonal" indicates that the characteristics of the antibody are obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies for use in accordance with the present invention can be prepared by a variety of techniques, including but not limited to hybridoma methods, recombinant DNA methods, phage display methods, and methods that utilize transgenic animals containing all or part of a human immunoglobulin locus, such methods and other exemplary methods for preparing monoclonal antibodies are described herein.

"naked antibody" refers to an antibody that is not conjugated to a heterologous moiety (e.g., a cytotoxic moiety) or radiolabeled. The naked antibody may be present in a pharmaceutical formulation.

"Natural antibody" refers to a naturally occurring immunoglobulin molecule having a different structure. For example, a native IgG antibody is a heterotetrameric glycoprotein of about 150,000 daltons, consisting of two identical light chains and two identical heavy chains that are disulfide-bonded. From N-terminus to C-terminus, each heavy chain has a variable region (VH), also known as a variable heavy or heavy chain variable domain, followed by three constant domains (CH 1, CH2 and CH 3). Similarly, each light chain has, from N-terminus to C-terminus, a variable region (VL), also known as a variable light chain domain or light chain variable domain, followed by a constant light Chain (CL) domain. The light chain of an antibody can be assigned to one of two types, called kappa (κ) and lambda (λ), based on the amino acid sequence of its constant domain.

The term "oligonucleotide" refers to a relatively short polynucleotide (e.g., less than about 250 nucleotides in length), including, but not limited to, single-stranded deoxyribonucleotides, single-or double-stranded ribonucleotides, RNA: DNA hybrids, and double-stranded DNA. Oligonucleotides, such as single-stranded DNA probe oligonucleotides, are typically synthesized by chemical methods, for example using commercially available automated oligonucleotide synthesizers. However, oligonucleotides can be prepared by a variety of other methods, including in vitro recombinant DNA-mediated techniques and by expression of DNA in cells and organisms.

The term "package insert" is used to refer to instructions typically included in commercial packaging for therapeutic products that contain information regarding the indications, usage, dosage, administration, combination therapy, contraindications, and/or warnings concerning the use of such therapeutic products.

The term "pharmaceutical formulation" refers to a formulation that is in a form that allows for the biological activity of the active ingredient contained therein to be effective, and that is free of additional components having unacceptable toxicity to the subject to which the formulation is to be administered.

By "pharmaceutically acceptable carrier" is meant a component of a pharmaceutical formulation that is not toxic to the subject except for the active ingredient. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, or preservatives.

The term "protein" as used herein, unless otherwise indicated, refers to any native protein from any vertebrate source, including mammals such as primates (e.g., humans) and rodents (e.g., mice and rats). The term includes "full-length" unprocessed protein, as well as any form of protein produced by processing in a cell. The term also encompasses naturally occurring protein variants, such as splice variants or allelic variants.

"percent (%) amino acid sequence identity" with respect to a reference polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical with amino acid residues in a reference polypeptide sequence, after aligning the candidate sequence with the reference polypeptide sequence and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and without regard to any conservative substitutions as part of the sequence identity. Alignments to determine percent amino acid sequence identity can be performed in a variety of ways within the skill in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, or Megalign (DNASTAR) software. One skilled in the art can determine appropriate parameters for aligning the sequences, including any algorithms necessary to achieve maximum alignment over the full length of the sequences being compared. However, for purposes herein, the sequence comparison computer program ALIGN-2 is used to generate values for% amino acid sequence identity. The ALIGN-2 sequence comparison computer program was written by Genentech, inc and the source code has been submitted with the user document to u.s.copy Office, washington d.c.,20559, where it was registered with us copyright registration number TXU 510087. The ALIGN-2 program is publicly available from Genettech, inc., south San Francisco, california, or may be compiled from this source code. The ALIGN-2 program should be compiled for use on a UNIX operating system, which includes the digital UNIX V4.0D. All sequence comparison parameters were set by the ALIGN-2 program and were unchanged.

In the case of amino acid sequence comparisons using ALIGN-2, the% amino acid sequence identity (which may alternatively be expressed as a percentage of the amino acid sequence identity of a given amino acid sequence A with or including a given amino acid sequence B) of a given amino acid sequence A to a given amino acid sequence B is calculated as follows:

100 times a fraction X/Y

Wherein X is the number of amino acid residues scored by the sequence alignment program ALIGN-2 as an identical match in an alignment of the program to A and B, and wherein Y is the total number of amino acid residues in B. It will be appreciated that where the length of amino acid sequence A is not equal to the length of amino acid sequence B, the% amino acid sequence identity of A to B will not be equal to the% amino acid sequence identity of B to A. Unless otherwise specifically indicated, all values of% amino acid sequence identity as used herein are obtained using the ALIGN-2 computer program as described in the preceding paragraph.

The terms "programmed death ligand 1" and "PD-L1" refer herein to native sequence PD-L1 polypeptides, polypeptide variants, and fragments of native sequence polypeptides and polypeptide variants (as further defined herein). The PD-L1 polypeptides described herein can be isolated from a variety of sources, such as from human tissue types or other sources, or made by recombinant or synthetic methods.

By "PD-L1 polypeptide variant" or variant thereof is meant a PD-L1 polypeptide (typically an active PD-L1 polypeptide) as defined herein that has at least about 80% amino acid sequence identity to any one of the native sequence PD-L1 polypeptide sequences as disclosed herein. Such PD-L1 polypeptide variants include, for example, PD-L1 polypeptides in which one or more amino acid residues are added or deleted at the N-terminus or C-terminus of the native amino acid sequence. Typically, a PD-L1 polypeptide variant will have at least about 80% amino acid sequence identity, or at least about 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% amino acid sequence identity, to a native sequence PD-L1 polypeptide sequence disclosed herein. Typically, a PD-L1 variant polypeptide is at least about 10 amino acids in length, alternatively at least about 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289 amino acids or more. Optionally, the PD-L1 variant polypeptide will have no more than one conservative amino acid substitution as compared to the native PD-L1 polypeptide sequence, or will contain no more than 2, 3, 4, 5, 6, 7, 8, 9, or 10 conservative amino acid substitutions as compared to the native PD-L1 polypeptide sequence.

A "native sequence PD-L1 polypeptide" comprises a polypeptide having the same amino acid sequence as a corresponding PD-L1 polypeptide derived from nature.

The term "PD-L1 axis binding antagonist" refers to a molecule that inhibits the interaction of a PD-L1 axis binding partner with its one or more binding partners to eliminate T cell dysfunction caused by signaling on the PD-1 signaling axis, resulting in restoration or enhancement of T cell function. As used herein, PD-L1 axis binding antagonists include PD-L1 binding antagonists and PD-1 binding antagonists as well as molecules that interfere with the interaction between PD-L1 and PD-1 (e.g., PD-L2-Fc fusions).

The term "PD-L1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, eliminates or interferes with signaling resulting from the interaction of PD-L1 with one or more of its binding partners (such as PD-1 or B7-1). In some embodiments, the PD-L1 binding antagonist is a molecule that inhibits the binding of PD-L1 to its binding partner. In particular aspects, the PD-L1 binding antagonist inhibits the binding of PD-L1 to PD-1 and/or B7-1. In some embodiments, PD-L1 binding antagonists include anti-PD-L1 antibodies, antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides, and others that reduce, block, inhibit, eliminate, or interfere with signaling resulting from the interaction of PD-L1 with one or more of its binding partners (such as PD-1 or B7-1). In one embodiment, the PD-L1 binding antagonist can reduce a negative costimulatory signal mediated by or through PD-L1 signaling mediated by a cell surface protein expressed on the T lymphocyte, thereby rendering the dysfunctional T cell less dysfunctional (e.g., increasing effector response to antigen recognition). In some embodiments, the PD-L1 binding antagonist is an anti-PD-L1 antibody. In a particular embodiment, the anti-PD-L1 antibody is atelizumab (CAS registry No. 1422185-06-5), also known as MPDL3280A, and as described herein. In another specific embodiment, the anti-PD-L1 antibody is yw243.55.S70 described herein. In another specific embodiment, the anti-PD-L1 antibody is MDX-1105 as described herein. In yet another specific aspect, the anti-PD-L1 antibody is MEDI4736 (dewalumab), as described herein. In yet another specific aspect, the anti-PD-L1 antibody is MSB0010718C (avizumab), as described herein.

As used herein, the term "PD-1 binding antagonist" refers to a molecule that reduces, blocks, inhibits, eliminates, or interferes with signaling resulting from the interaction of PD-1 with one or more of its binding partners (such as PD-L1 and/or PD-L2). In some embodiments, the PD-1 binding antagonist is a molecule that inhibits the binding of PD-1 to its binding partner. In particular aspects, the PD-1 binding antagonist inhibits the binding of PD-1 to PD-L1 and/or PD-L2. For example, PD-1 binding antagonists include anti-PD-1 antibodies and antigen-binding fragments thereof, immunoadhesins, fusion proteins, oligopeptides, small molecule antagonists, polynucleotide antagonists, and others that reduce, block, inhibit, eliminate, or otherwise interfere with signaling resulting from the interaction of PD-1 with PD-L1 and/or PD-L2. In one embodiment, the PD-1 binding antagonist reduces negative signals mediated (by PD-1 or PD-L1 mediated signaling) by or through cell surface proteins expressed on T lymphocytes or other cells, thereby resulting in reduced dysfunctional T cell dysfunction. In some embodiments, the PD-1 binding antagonist is an anti-PD-1 antibody. In a particular aspect, the PD-1 binding antagonist is MDX-1106 (nivolumab). In another specific aspect, the PD-1 binding antagonist is MK-3475 (pembrolizumab). In another specific aspect, the PD-1 binding antagonist is MEDI-0680 (AMP-514). In another specific aspect, the PD-1 binding antagonist is PDR001. In another specific aspect, the PD-1 binding antagonist is REGN2810. In another particular aspect, the PD-1 binding antagonist is BGB-108. In another specific aspect, the PD-1 binding antagonist is AMP-224.

"Polynucleotide" or "nucleic acid" are used interchangeably herein to refer to a polymer of nucleotides of any length and include DNA and RNA. A nucleotide may be a deoxyribonucleotide, a ribonucleotide, a modified nucleotide or base, and/or an analog thereof, or any substrate that can be incorporated into a polymer by a DNA or RNA polymerase or by a synthetic reaction. Polynucleotides may comprise modified nucleotides, such as methylated nucleotides and analogs thereof. If present, the nucleotide structure may be modified before or after assembly of the polymer. The sequence of nucleotides may be interrupted by non-nucleotide components. The polynucleotide may be further modified after synthesis, for example by conjugation with a label. Other types of modifications include, for example, "capping," replacing one or more of the naturally occurring nucleotides with similar internucleotide modifications, such as, for example, those having uncharged bonds (e.g., methyl phosphates, phosphotriesters, phosphoramidates, carbamates) and having charged bonds (e.g., phosphorothioates, phosphorodithioates, etc.), those containing pendant groups, such as, for example, proteins (e.g., nucleases, toxins, antibodies, signal peptides, ply-L-lysine, etc.), those containing intercalators (e.g., acridine, psoralen, etc.), those containing chelators (e.g., metals, radioactive metals, boron, oxidative metals, etc.), those containing alkylating agents, those having modified bonds (e.g., alpha anomeric nucleic acids, etc.), and unmodified forms of the polynucleotide. In addition, any hydroxyl group typically present in a sugar may be replaced (e.g., phosphate group), protected by a standard protecting group, or activated to make additional linkages to additional nucleotides, or may be conjugated to a solid or semi-solid support. The 5 'and 3' terminal OH groups may be phosphorylated or partially substituted with an amine or organic end-capping group of 1-20 carbon atoms. Other hydroxyl groups may also be derivatized as standard protecting groups. Polynucleotides may also comprise similar forms of ribose or deoxyribose as are generally known in the art, including, for example, 2 '-O-methyl-, 2' -O-allyl, 2 '-fluoro-or 2' -azido-ribose, carbocyclic sugar analogs, α -anomeric sugars, epimeric sugars (such as arabinose, xylose, or lyxose, pyranose, furanose, heptaheptose), acyclic analogs, and abasic nucleoside analogs such as methyl riboside. One or more phosphodiester linkages may be replaced by alternative linking groups. These alternative linking groups include, but are not limited to, embodiments in which the phosphate is replaced by P (O) S ("thioester"), P (S) S ("dithioate"), (O) NR2 ("amidate"), P (O) R, P (O) OR ', CO, OR CH2 ("methylal"), where each R OR R' is independently H OR a substituted OR unsubstituted alkyl (1-20C) optionally containing an ether (-O-) bond, aryl, alkenyl, cycloalkyl, cycloalkenyl, OR aralkyl (araldyl). Not all linkages in a polynucleotide need be identical. The foregoing description applies to all polynucleotides referred to herein, including RNA and DNA.

As used herein, "polymerase chain reaction" or "PCR" techniques generally refer to the procedure for amplifying minute quantities of a particular nucleic acid, RNA and/or DNA fragment as described in U.S. patent No. 4,683,195, issued on 28/7/1987. Generally, it is desirable to obtain sequence information from the end of the region of interest or from regions other than the end so that oligonucleotide primers can be designed; these primers are identical or similar in sequence to opposite strands of the template to be amplified. The 5' terminal nucleotides of the two primers may coincide with the ends of the amplified material. PCR can be used to amplify specific RNA sequences and specific DNA sequences from total genomic DNA, as well as cDNA transcribed from total cellular RNA, phage or plasmid sequences, and the like. See generally Mullis et al, cold Spring Harbor symp. Quant.biol.,51 (1987); erlich, eds, PCR Technology (Stockton Press, NY, 1989). As used herein, PCR is considered to be one, but not the only example, of a nucleic acid polymerase reaction method for amplifying a nucleic acid test sample, including the use of known nucleic acids (DNA or RNA) as primers and the use of a nucleic acid polymerase to amplify or generate a particular nucleic acid fragment, or to amplify or generate a particular nucleic acid fragment that is complementary to a particular nucleic acid.

As used herein, the term "reverse transcriptase polymerase chain reaction" or "RT-PCR" refers to the replication and amplification of RNA sequences. In this method, for example, reverse transcription is coupled to PCR as described in U.S. patent No. 5,322,770, which is incorporated by reference herein in its entirety. In RT-PCR, an RNA template is converted to cDNA due to the reverse transcriptase activity of the enzyme, and then amplified using the polymerization activity of the same or a different enzyme. Thermostable and thermolabile reverse transcriptases and polymerases may be used. "reverse transcriptase" (RT) may include reverse transcriptases from retroviruses, other viruses, and DNA polymerases that exhibit reverse transcriptase activity.

As used herein, the term "reverse transcriptase quantitative polymerase chain reaction" or "RT-qPCR" is a form of PCR in which the nucleic acid to be amplified is RNA that is first reverse transcribed into cDNA, and the amount of PCR product is measured in each step of the PCR reaction.

"quantitative real-time polymerase chain reaction" or "qRT-PCR" refers to a form of PCR in which the amount of PCR product is measured in each step of the PCR reaction. In the classes including Cronin et al, am.J.Pathol.164 (1): 35-42 (2004); and Ma et al, cancer Cell 5, 607-616 (2004).

The term "multiplex PCR" refers to a single PCR reaction performed on nucleic acids obtained from a single source (e.g., an individual) using more than one primer set, with the aim of amplifying two or more DNA sequences in a single reaction.

The term "RNA-seq," also known as "Whole Transcriptome Shotgun Sequencing (WTSS)," refers to sequencing and/or quantifying cDNA using high-throughput sequencing techniques to obtain information about the RNA content of a sample. Publications describing RNA-seq include: wang et al, nature Reviews Genetics 10 (1): 57-63 (2009); ryan et al, bioTechniques 45 (1): 81-94 (2008); and Maher et al, nature 458 (7234): 97-101 (2009).

The term "polynucleotide" when used in the singular or plural generally refers to any polyribonucleotide or polydeoxyribonucleotide, which may be unmodified RNA or DNA or modified RNA or DNA. Thus, for example, polynucleotides as defined herein include, but are not limited to: single-stranded and double-stranded DNA; DNA comprising single-stranded and double-stranded regions; single-and double-stranded RNA; RNA comprising single-stranded and double-stranded regions; and hybrid molecules comprising DNA and RNA (which may be single-stranded, or more typically double-stranded, or comprise single-and double-stranded regions). Furthermore, the term "polynucleotide" as used herein refers to a triple-stranded region comprising RNA or DNA or both RNA and DNA. The chains in such regions may be from the same molecule or from different molecules. A region may comprise all of one or more of the molecules, but typically comprises only one region of a portion of the molecule. One of the molecules having a triple-helical region is typically an oligonucleotide. The term "polynucleotide" specifically includes cDNA. The term includes DNA (including cDNA) and RNA that contain one or more modified bases. Thus, a DNA or RNA having a backbone modified for stability or other reasons is a "polynucleotide" as described herein. In addition, the term "polynucleotide" as defined herein includes DNA or RNA containing unusual bases (such as inosine) or modified bases (such as tritiated bases). In general, the term "polynucleotide" encompasses all chemically, enzymatically and/or metabolically modified forms of unmodified polynucleotides, as well as chemical forms of DNA and RNA that are characteristic of viruses and cells, including simple and complex cells.

Any endpoint that is indicated to be beneficial to an individual can be used to assess "response to treatment," "responsiveness to treatment," or "benefit from treatment," including but not limited to (1) inhibition of disease progression (e.g., cancer progression) to some extent, including slowing and complete arrest; (2) reducing tumor size; (3) Inhibit (i.e., reduce, slow, or completely stop) cancer cell infiltration into adjacent peripheral organs and/or tissues; (4) Inhibit (i.e., reduce, slow, or completely stop) metastasis; (5) Alleviating, to some extent, one or more symptoms associated with the disease or condition (e.g., cancer); (6) Increased or prolonged survival, including overall survival (OS HR < 1) and progression-free survival (PFS HR < 1); and/or (9) reduce mortality at a given time point following treatment (e.g., treatment comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)). As used herein, a patient that is "non-responsive" to a particular form of treatment refers to a patient that fails to exhibit any or all of the above-described benefits after administration of a targeted therapy.

As used herein, "progression-free survival" or "PFS" refers to the length of time during and after treatment during which the disease being treated (e.g., cancer, such as lung cancer (e.g., NSCLC, including non-squamous NSCLC and squamous NSCLC), bladder cancer (e.g., UC), renal cancer (e.g., RCC), or breast cancer (e.g., TNBC)) does not progress or worsen. Progression-free survival can include the amount of time an individual experiences a complete response or a partial response, as well as the amount of time an individual experiences stable disease.

As used herein, "overall survival" or "OS" refers to the length of time a subject is likely to survive during and after treatment for a particular period of time (e.g., 6 months, 1 year, 2 years, 3 years, 4 years, 5 years, 10 years, 15 years, 20 years, or more than 20 years from diagnosis or treatment initiation).

As used herein, "complete response" or "CR" refers to the disappearance of all signs of cancer in response to treatment. This does not necessarily mean that the cancer has already cured.

As used herein, "partial response" or "PR" refers to a reduction in the size of one or more tumors or lesions or a reduction in the extent of cancer in vivo in response to treatment.

As used herein, "risk ratio" or "HR" is a statistical definition of the rate of occurrence of an event. For the purposes of the present invention, a risk ratio is defined as representing the probability of an event (e.g., PFS or OS) in an experimental (e.g., treatment) group (group)/cohort (arm) divided by the probability of an event in a control group (group)/cohort (arm) at any particular point in time. An HR value of 1 indicates that the relative risk (e.g., death) of the endpoint is equal in both the "treatment" group and the "control" group; a value greater than 1 indicates a greater risk in the treated group relative to the control group; a value less than 1 indicates a greater risk for the control group relative to the treated group. The "risk ratio" in the progression-free survival analysis (i.e. PFS HR) summarizes the difference between the two progression-free survival curves, representing a reduced risk of mortality for the treatment compared to the control over the follow-up period. The "risk ratio" in the overall survival analysis (i.e. OS HR) summarizes the difference between the two overall survival curves, representing a reduced risk of mortality for the treatment compared to the control over the follow-up period.

By "extended survival" or "extension of survival" is meant an increase in the overall survival or progression-free survival of a treated individual relative to an untreated individual (i.e., relative to an individual not treated with a drug) or relative to an individual not expressing a specified level of a biomarker and/or relative to an individual treated with an approved anti-neoplastic agent. Objective response refers to a measurable response, including Complete Response (CR) or Partial Response (PR).

By "reduce or inhibit" is meant the ability to cause an overall decrease of 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95% or more. Reduced or inhibited can refer to the symptoms of a condition being treated (e.g., cancer, such as lung cancer (e.g., NSCLC, including non-squamous NSCLC and squamous NSCLC), bladder cancer (e.g., UC), renal cancer (e.g., RCC), or breast cancer (e.g., TNBC)), the presence or size of a metastatic tumor, or the size of a primary tumor.

As used herein, "reference sample," "reference cell," "reference tissue," "control sample," "control cell," or "control tissue" refers to a sample, cell, tissue, standard, or level for purposes of comparison. In one embodiment, the reference sample, reference cell, reference tissue, control sample, control cell, or control tissue is obtained from the same subject or individual. In another embodiment, the reference sample is obtained from one or more individuals that are not subjects or individuals. In any of the preceding embodiments, one or more individuals from whom the reference sample, reference cell, reference tissue, control sample, control cell, or control tissue is obtained have cancer. In certain embodiments, one or more individuals from whom the reference sample, reference cells, reference tissue, control sample, control cells, or control tissue is obtained have cancer and have been previously treated with an anti-cancer therapy (e.g., one or more doses of a PD-L1 axis binding antagonist). In other embodiments, one or more individuals from whom the reference sample, reference cell, reference tissue, control sample, control cell, or control tissue is obtained have cancer and are untreated. In any of the preceding embodiments, the subject/individual and one or more individuals that are not the subject or individual have the same cancer. In yet another embodiment, the reference sample, reference cell, reference tissue, control sample, control cell, or control tissue is obtained from a healthy and/or non-diseased body part (e.g., tissue or cell) of the same subject or individual. For example, healthy and/or non-diseased cells or tissues adjacent to a diseased cell or tissue (e.g., cells or tissues adjacent to a tumor). In another embodiment, the reference sample is obtained from untreated tissue and/or cells of the body of the same subject or individual. In yet another embodiment, the reference sample, reference cell, reference tissue, control sample, control cell, or control tissue is obtained from a healthy and/or non-diseased portion (e.g., tissue or cell) of the body of an individual that is not the subject or individual. In yet another embodiment, the reference sample, reference cell, reference tissue, control sample, control cell, or control tissue is obtained from untreated tissue and/or cells that are not part of the individual's body of the subject or individual.

As used herein, the term "sample" refers to a composition obtained or derived from a subject and/or individual of interest that comprises, for example, cells and/or other molecular entities to be characterized and/or identified based on physical, biochemical, chemical, and/or physiological characteristics. For example, the phrase "disease sample" and variations thereof refers to any sample obtained from a subject of interest that is expected or known to comprise the cellular and/or molecular entities to be characterized. Samples include, but are not limited to, primary or cultured cells or cell lines, cell supernatants, cell lysates, platelets, serum, plasma, vitreous humor, lymph fluid, synovial fluid, follicular fluid, semen, amniotic fluid, milk, whole blood, blood-derived cells, urine, cerebrospinal fluid, saliva, sputum, tears, sweat, mucus, tumor lysate and tissue culture medium, tissue extracts such as homogenized tissue, tumor tissue, cell extracts, and combinations thereof.

As used herein, the terms "individual," "patient," and "subject" are used interchangeably and refer to any single animal, more preferably a mammal (including, e.g., non-human animals, e.g., cats, dogs, horses, rabbits, zoo animals, cows, pigs, sheep, and non-human primates), in need of treatment. In certain embodiments, the individual, patient, or subject is a human.

As used herein, "treatment" (and grammatical variations thereof, such as "treatment" or "treating") refers to a clinical intervention that attempts to alter the natural course of the subject to be treated, and may be for the purpose of prophylaxis or in the course of clinical pathology. Desirable therapeutic effects include, but are not limited to, preventing the onset or recurrence of a disease (e.g., cancer, such as lung cancer (e.g., NSCLC, including non-squamous NSCLC and squamous NSCLC), bladder cancer (e.g., UC), renal cancer (e.g., RCC), or breast cancer (e.g., TNBC)), alleviating symptoms, alleviating any direct or indirect pathological consequences of the disease, preventing metastasis, reducing the rate of disease progression, improving or mitigating a condition, and alleviating or improving prognosis. In some embodiments, the treatment described herein is used to delay the progression or slow the progression of a disease (e.g., cancer, such as lung cancer (e.g., NSCLC, including non-squamous NSCLC and squamous NSCLC), bladder cancer (e.g., UC), renal cancer (e.g., RCC), or breast cancer (e.g., TNBC)). In some cases, treatment may increase Overall Survival (OS) (e.g., by about 20% or more, about 25% or more, about 30% or more, about 35% or more, about 40% or more, about 45% or more, about 50% or more, about 55% or more, about 60% or more, about 65% or more, about 70% or more, about 75% or more, about 80% or more, about 85% or more, about 90% or more, about 95% or more, about 96% or more, about 97% or more, about 98% or more, or about 99% or more). In some cases, treatment may increase OS, e.g., by about 5% to about 500%, e.g., about 10% to about 450%, e.g., about 20% to about 400%, e.g., about 25% to about 350%, e.g., about 30% to about 400%, e.g., about 35% to about 350%, e.g., about 40% to about 300%, e.g., about 45% to about 250%, e.g., about 50% to about 200%, e.g., about 55% to about 150%, e.g., about 60% to about 100%, e.g., about 65% to about 100%, e.g., about 70% to about 100%, e.g., about 75% to about 100%, e.g., about 80% to about 100%, e.g., about 85% to about 100%, e.g., about 90% to about 100%, e.g., about 95% to about 100%, e.g., about 98% to about 100%. In some cases, treatment may increase progression-free survival (PFS) (e.g., an increase of about 20% or more, about 25% or more, about 30% or more, about 35% or more, about 40% or more, about 45% or more, about 50% or more, about 55% or more, about 60% or more, about 65% or more, about 70% or more, about 75% or more, about 80% or more, about 85% or more, about 90% or more, about 95% or more, about 96% or more, about 97% or more, about 98% or more, or about 99% or more). In some cases, treatment may increase PFS, e.g., by about 5% to about 500%, e.g., about 10% to about 450%, e.g., about 20% to about 400%, e.g., about 25% to about 350%, e.g., about 30% to about 400%, e.g., about 35% to about 350%, e.g., about 40% to about 300%, e.g., about 45% to about 250%, e.g., about 50% to about 200%, e.g., about 55% to about 150%, e.g., about 60% to about 100%, e.g., about 65% to about 100%, e.g., about 70% to about 100%, e.g., about 75% to about 100%, e.g., about 80% to about 100%, e.g., about 85% to about 100%, e.g., about 90% to about 100%, e.g., about 95% to about 100%, e.g., about 98% to about 100%.

"tissue sample" or "cell sample" refers to a collection of similar cells obtained from a tissue of a subject or individual. The source of the tissue or cell sample may be solid tissue from fresh, frozen and/or preserved organs, tissue samples, biopsies and/or aspirates; blood or any blood component, such as plasma; body fluids, such as cerebrospinal fluid, amniotic fluid, peritoneal fluid, or interstitial fluid; cells at any time during pregnancy or development of the subject. The tissue sample may also be primary or cultured cells or cell lines. Optionally, the tissue or cell sample is obtained from a disease (e.g., cancer, e.g., lung cancer (e.g., NSCLC, including non-squamous NSCLC and squamous NSCLC), bladder cancer (e.g., UC), renal cancer (e.g., RCC), or breast cancer (e.g., TNBC)) tissue/organ. Tissue samples may contain compounds that do not naturally mix with tissue in the natural environment, such as preservatives, anticoagulants, buffers, fixatives, nutrients, antibiotics, and the like.

For purposes herein, a "section" of a tissue sample refers to a single portion or piece of the tissue sample, e.g., a thin slice of tissue or cells cut from the tissue sample. It is understood that multiple sections of a tissue sample may be taken and analyzed, provided that it is understood that the same section of a tissue sample may be analyzed at the morphological and molecular level, or may be analyzed for polypeptides and polynucleotides.

The terms "tertiary lymphoid structure" and "TLS" are used interchangeably herein and refer to ectopic lymphoid structures that may form in non-lymphoid tissues (e.g., at sites of chronic inflammation, including tumors). These terms may refer to structures of different tissues, for example, from lymphocyte clusters to isolated structures reminiscent of secondary lymphoid organs. For example, these terms encompass TLS-like structures. In some cases, TLS may comprise distinct T-cell and B-cell compartments, a Fibroblast Reticulocyte (FRC) network, peripheral lymph node addressin (PNAd) ⁺ ) Evidence of High Endothelial Venules (HEV), follicular Dendritic Cells (FDC), class switching and reactive Germinal Centers (GC) in the B cell region and/or activation-induced expression of cytidine deaminase (AID), an enzyme expressed in GC B cells involved in somatic hypermutation initiation and immunoglobulin gene class switching. For a review of TLS in cancer, see Colbeck et al front.Immunol.8:1830,2017.

As used herein, the term "tumor" refers to all neoplastic cell growth and proliferation, whether malignant or benign, and all pre-cancerous and cancerous cells and tissues. The terms "cancer," "cancerous," "cell proliferative disorder," "proliferative disorder," and "tumor" are not mutually exclusive herein.

The term "variable region" or "variable domain" refers to the domain of an antibody heavy or light chain that is involved in binding of the antibody to an antigen. The variable domains of the heavy and light chains of natural antibodies (VH and VL, respectively) generally have a similar structure, with each domain containing four conserved Framework Regions (FR) and three hypervariable regions (HVRs). (see, e.g., kindt et al, kuby Immunology, 6 th edition, w.h.freeman and co., page 91 (2007)) a single VH or VL domain may be sufficient to confer antigen binding specificity. Furthermore, antibodies that bind a particular antigen can be isolated using the VH or VL domains, respectively, from antibodies that bind the antigen to screen libraries of complementary VL or VH domains. See, e.g., portolano et al, J.Immunol.150:880-887 (1993); clarkson et al, nature 352, 624-628 (1991).

Diagnostic method

Provided herein are methods for identifying individuals having cancer (e.g., lung cancer (e.g., non-small cell lung cancer (NSCLC)), bladder cancer (e.g., urothelial Cancer (UC)), kidney cancer (e.g., renal Cell Carcinoma (RCC)) or breast cancer (e.g., triple Negative Breast Cancer (TNBC)) that may benefit from treatment comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., an atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

Further provided herein are methods for selecting a therapy for an individual having cancer (e.g., lung cancer (e.g., NSCLC), bladder cancer (e.g., UC), renal cancer (e.g., RCC), or breast cancer (e.g., TNBC)); a method for determining whether an individual having cancer is likely to respond to treatment comprising a PD-L1 axis binding antagonist; a method for predicting responsiveness of an individual having cancer to a binding antagonist comprising the PD-L1 axis; and methods of monitoring the response of an individual having cancer to a treatment regimen comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

Any of the methods provided herein can comprise determining the presence and/or expression level of any of the biomarkers disclosed herein. For example, a biomarker may comprise the presence and/or level of expression of a biomarker listed in any one of tables 1 to 17 below in a sample obtained from an individual; the presence of TLS in a sample obtained from an individual; the number of B cells in a sample obtained from an individual; the case of clonally expanded B cells in a sample from an individual; and/or combinations thereof. Any suitable sample may be used, for example, any sample type disclosed herein, including tumor samples.

Any of the methods provided herein can further comprise selecting a therapy for the individual, such as a therapy comprising a PD-L1 axis binding antagonist (e.g., as described below in section IV).

Any of the methods provided herein can further comprise administering to the individual a separate PD-L1 axis binding antagonist (e.g., as described in section IV below).

Table 1: exemplary biomarkers

CD79A
	SLAMF7
BTK
	TNFRSF17
IGJ
	IGLL5
RBPJ
	MZB1
CCL2
	CCL3
CCL4
	CCL5
CCL8
	CCL18
CCL19
	CCL21
CXCL9
	CXCL10
CXCL11
	CXCL13

For example, provided herein is a method of identifying an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) who is likely to benefit from treatment comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attrituzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), the method comprising determining, in a sample from the individual, the expression levels of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) genes listed in table 1, wherein the individual is likely to benefit from identification of the PD axis binding antagonist in the event that the immune score expression level of the one or more genes is higher than a reference immune expression level of the one or more genes.

In another example, provided herein is a method of selecting a therapy for an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising determining the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) of the genes listed in table 1 in a sample from the individual, wherein the individual is identified as likely to benefit from treatment with a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attritol antagonist) or a PD-1 binding antagonist (e.g., an anti-PD 1 antibody)), where the immune score expression level of the one or more genes is higher than a reference immune score expression level of the one or more genes.

In some cases, the immune score expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) genes in the sample is greater than the reference immune score expression level, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attritumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)). Any suitable PD-L1 axis binding antagonist can be administered, e.g., any PD-L1 axis binding antagonist provided herein (e.g., as described in section IV below).

Any suitable immuno-score reference expression level can be used. In some cases, the immune score reference expression level is an immune score expression level of one or more genes in a reference population. In some cases, the reference population is a population of individuals with cancer. In some cases, the population of individuals includes a first subset of individuals who have received treatment with a PD-L1 axis binding antagonist and a second subset of individuals who have received treatment with a therapy that does not contain a PD-L1 axis binding antagonist. In some cases, the reference immune score expression level visibly differentiates each of the first subset of individuals from the second subset of individuals based on a significant difference between the individual's responsiveness to treatment with a PD-L1 axis binding antagonist and the individual's responsiveness to treatment with a therapy that does not contain a PD-L1 axis binding antagonist, above the reference immune score expression level, wherein the individual's responsiveness to treatment with a PD-L1 axis binding antagonist is significantly improved relative to the individual's responsiveness to treatment with a therapy that does not contain a PD-L1 axis binding antagonist. In some cases, the therapy without the PD-L1 axis binding antagonist comprises an anti-neoplastic agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, or a combination thereof. In some cases, the therapy without the PD-L1 axis binding antagonist comprises a chemotherapeutic agent. In some cases, the chemotherapeutic agent is docetaxel. In some cases, responsiveness to treatment comprises prolongation of OS, prolongation of progression-free survival (PFS), or an increase in the determined optimal overall response (BCOR). In some cases, responsiveness to treatment includes an extension of Overall Survival (OS). In some cases, the reference immune score expression level is the median of the expression levels of each of the one or more genes in the reference population. In some cases, the median expression level is the median of the mean Z scores for the expression levels of each of the one or more genes in the reference population.

Any suitable sample may be used. In some cases, the sample is a tissue sample, a cell sample, a whole blood sample, a plasma sample, a serum sample, or a combination thereof. In some cases, the tissue sample is a tumor tissue sample. In some cases, the tumor sample is a tumor tissue sample. In some cases, the tumor tissue sample comprises tumor cells, tumor infiltrating immune cells, stromal cells, NAT cells, or a combination thereof. In some cases, the tumor tissue sample is a formalin-fixed and paraffin-embedded (FFPE) sample, an archived sample, a fresh sample, or a frozen sample. In some cases, the tumor tissue sample is an FFPE sample.

The cancer may be any suitable type of cancer. In some cases, the cancer is lung cancer, kidney cancer, bladder cancer, breast cancer, colorectal cancer, ovarian cancer, pancreatic cancer, stomach cancer, esophageal cancer, mesothelioma, melanoma, head and neck cancer, thyroid cancer, sarcoma, prostate cancer, glioblastoma, cervical cancer, thymus cancer, leukemia, lymphoma, myeloma, mycosis fungoides, merkel cell carcinoma, or a hematological malignancy. In some cases, the cancer is lung, kidney, bladder, or breast cancer. In some cases, the lung cancer is non-small cell lung cancer (NSCLC). In some cases, the NSCLC is non-squamous NSCLC. In some cases, the NSCLC is squamous NSCLC.

In some cases, the benefit comprises a prolongation of OS in the individual, a prolongation of PFS in the individual, and/or an improvement in BCOR in the individual as compared to treatment without the PD-L1 axis binding antagonist. In some cases, the benefit includes a prolongation of OS in the individual as compared to treatment without the PD-L1 axis binding antagonist.

A.B cell characteristics

(i) Gene signature associated with B cells

In some aspects, the methods provided herein can involve determining the expression level of one or more genes in a B cell signature. Any suitable B cell characteristics may be used. For example, a B cell characteristic can include one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) genes listed in table 2.

Table 2: exemplary B cell signature genes

CD79A
	CD19
BANK1
	JCHAIN
SLAMF7
	BTK
TNFRSF17
	IGJ
IGLL5
	RBPJ
MZB1

For example, provided herein is a method of identifying an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) who is likely to benefit from treatment comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attrituzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), the method comprising determining, in a sample from the individual, expression levels of one or more of genes CD79A, CD19, BANK1, jcain, SLAMF7, BTK TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) where an individual who is likely to benefit from identification of an immune expression level of the one or more genes that is higher than an immune expression level of the PD-L1 axis binding antagonist.

In another example, provided herein is a method of selecting a therapy for an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising determining the expression level of one or more of genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) in a sample from the individual, wherein the individual is identified as likely to benefit from treatment of the individual with a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 binding antibody, e.g., attritol 1-PD-binding antibody) or an anti-PD 1-bead antibody, e.g., anti-PD 1 monoclonal antibody).

In some cases, the method comprises determining the expression level of one of genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB 1.

In some cases, the method comprises determining the expression level of CD 79A.

In some cases, the method comprises determining the expression level of CD 19.

In some cases, the method comprises determining an expression level of BANK 1.

In some cases, the method comprises determining the expression level of JCHAIN.

In some cases, the method comprises determining the expression level of SLAMF 7.

In some cases, the method includes determining an expression level of BTK.

In some cases, the method comprises determining the expression level of TNFRSF 17.

In some cases, the method comprises determining the expression level of IGJ.

In some cases, the method includes determining the expression level of IGLL 5.

In some cases, the method comprises determining an expression level of RBPJ.

In some cases, the method comprises determining an expression level of MZB 1.

In some cases, the immune score expression level of one or more genes in the sample is greater than the reference immune score expression level, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)). Any suitable PD-L1 axis binding antagonist can be administered, e.g., any PD-L1 axis binding antagonist provided herein (e.g., as described in section IV below).

Any suitable immuno-score reference expression level can be used. In some cases, the immune score reference expression level is an immune score expression level of one or more genes in the reference population. In some cases, the reference population is a population of individuals with cancer. In some cases, the population of individuals includes a first subset of individuals who have received treatment with a PD-L1 axis binding antagonist and a second subset of individuals who have received treatment with a therapy that does not contain a PD-L1 axis binding antagonist. In some cases, the reference immune score expression level visibly differentiates each of the first subset of individuals from the second subset of individuals based on a significant difference between the individual's responsiveness to treatment with a PD-L1 axis binding antagonist and the individual's responsiveness to treatment with a therapy that does not contain a PD-L1 axis binding antagonist, above the reference immune score expression level, wherein the individual's responsiveness to treatment with a PD-L1 axis binding antagonist is significantly improved relative to the individual's responsiveness to treatment with a therapy that does not contain a PD-L1 axis binding antagonist. In some cases, the therapy without the PD-L1 axis binding antagonist comprises an anti-neoplastic agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, or a combination thereof. In some cases, the therapy without the PD-L1 axis binding antagonist comprises a chemotherapeutic agent. In some cases, the chemotherapeutic agent is docetaxel. In some cases, responsiveness to treatment comprises prolongation of OS, prolongation of PFS, or an increase in BCOR. In some cases, responsiveness to treatment includes an extension of Overall Survival (OS). In some cases, the reference immune score expression level is the median of the expression levels of each of the one or more genes in the reference population. In some cases, the median expression level is the median of the mean Z scores for the expression levels of each of the one or more genes in the reference population.

For example, provided herein is a method of identifying an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) who may benefit from treatment comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attrituzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), the method comprising determining the expression levels of one or more of genes CD79A, CD19, BANK1, jcain, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) in a sample from the individual, wherein an individual who has a higher immune expression level than the reference gene or genes whose immune expression level would benefit from treatment comprises an extended PD-axis binding to the individual, wherein the subject's PD-1 would benefit from treatment using the antagonist.

In another example, provided herein is a method of selecting a therapy for an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising determining the expression level of one or more of genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, rbj, and MZB1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) in a sample from the individual, wherein the individual is identified as likely to benefit from treatment with a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 binding antagonist, such as an attritol antibody) or an anti-PD 1 binding antibody, wherein the treatment does not include treatment of the individual with the PD-1 axis binding antagonist, such as an anti-PD 1 binding antagonist (e.g., an anti-PD 1) antibody, wherein the individual does not benefit from treatment with the anti-PD 1, such as an anti-PD-OS antagonist.

In some cases, the immune score expression level of one or more genes in the sample is higher than the reference immune score expression level, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)). Any suitable PD-L1 axis binding antagonist can be administered, e.g., any PD-L1 axis binding antagonist provided herein (e.g., as described in section IV below).

In some cases, the genes include two or more of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB 1.

For example, provided herein is a method of identifying an individual having cancer who may benefit from treatment comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), the method comprising determining the expression level of two or more of genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) in a sample from the individual, wherein the individual is identified as a treated individual who may comprise a PD-L1 axis binding antagonist if the immune score expression level of the two or more genes is higher than the reference immune score expression level of the two or more genes.

In another example, provided herein is a method of selecting a therapy for an individual having cancer, the method comprising determining the expression level of two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein the individual is identified as an individual likely to benefit from treatment comprising a PD-L1 axis binding antagonist if the immune score expression level of the two or more genes is greater than a reference immune score expression level of the two or more genes.

In another example, provided herein is a method of identifying an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) who may benefit from treatment with a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attrituximab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) comprising determining expression levels of two or more of genes CD79A, CD19, BANK1, jcain, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) in a sample from the individual, wherein an immune expression level of the two or more genes is higher than an immune expression level of the two or more genes, wherein the identification of the PD-L axis binding antagonist is included in the individual who may benefit from treatment with prolonged PD-1 axis binding.

In yet another example, provided herein is a method of selecting a therapy for an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising determining the expression level of two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) of genes CD79A, CD19, BANK1, jcain, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, rbj, and MZB1 in a sample from the individual, wherein the individual is identified as likely to benefit from treatment with a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antagonist, e.g., attritrin) or an anti-PD 1 binding antagonist, wherein the treatment of the individual does not comprise prolonged PD-1 axis binding of the PD-OS treatment.

In some cases, the immune score expression level of two or more genes in the sample is higher than the reference immune score expression level, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)). Any suitable PD-L1 axis binding antagonist can be administered, e.g., any PD-L1 axis binding antagonist provided herein (e.g., as described in section IV below).

Any combination of B cell signature genes can be determined. For example, the combination may include two genes selected from CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1, e.g., any one of the combinations listed in table 3. In another example, the combination can include three genes selected from CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1, e.g., any one of the combinations listed in table 4. In another example, the combination can include four genes selected from CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1, e.g., any one of the combinations listed in table 5. In another example, the combination may include five genes selected from CD79A, CD19, BANK1, jchan, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1, e.g., any one of the combinations listed in table 6. In another example, the combination can include six genes selected from CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1, e.g., any one of the combinations listed in table 7. In another example, the combination may include seven genes selected from CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1, e.g., any one of the combinations listed in table 8.

Table 3: exemplary two-Gene combinations of B-cell signature genes

Table 4: exemplary three-Gene combinations of B-cell signature genes

Table 5: exemplary four Gene combinations of B cell signature genes

Table 6: exemplary five-Gene combinations of B-cell signature genes

Table 7: exemplary six-Gene combinations of B cell signature genes

Table 8: exemplary seven Gene combinations of B cell signature genes

CD79A, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, and RBPJ
	CD79A, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, and MZB1
CD79A, SLAMF7, BTK, TNFRSF17, IGJ, RBPJ, and MZB1
	CD79A, SLAMF7, BTK, TNFRSF17, IGLL5, RBPJ, and MZB1
CD79A, SLAMF7, BTK, IGJ, IGLL5, RBPJ, and MZB1
	CD79A, SLAMF7, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1
CD79A, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1
	SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1
CD19, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, and RBPJ
	CD19, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, and MZB1
CD19, SLAMF7, BTK, TNFRSF17, IGJ, RBPJ, and MZB1
	CD19, SLAMF7, BTK, TNFRSF17, IGLL5, RBPJ, and MZB1
CD19, SLAMF7, BTK, IGJ, IGLL5, RBPJ, and MZB1
	CD19, SLAMF7, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1
CD19, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1
	BANK1, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, and RBPJ
BANK1, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, and MZB1
	BANK1, SLAMF7, BTK, TNFRSF17, IGJ, RBPJ, and MZB1
BANK1, SLAMF7, BTK, TNFRSF17, IGLL5, RBPJ, and MZB1
	BANK1, SLAMF7, BTK, IGJ, IGLL5, RBPJ, and MZB1
BANK1, SLAMF7, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1
	BANK1, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1
JCHAIN、SLAMF7、BTK、TNFRSF17、IGJ、IGLL5 and RBPJ
	JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, and MZB1
JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, RBPJ, and MZB1
	JCHAIN, SLAMF7, BTK, TNFRSF17, IGLL5, RBPJ, and MZB1
JCHAIN, SLAMF7, BTK, IGJ, IGLL5, RBPJ, and MZB1
	JCHAIN, SLAMF7, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1
JCHAIN, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1

In some cases, the genes include CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1.

In some cases, the expression level is a nucleic acid expression level. For example, in some cases, the nucleic acid expression level is an mRNA expression level. mRNA expression levels can be determined using any suitable method (e.g., any method disclosed herein). In some cases, mRNA expression levels are determined by RNA-seq, RT-qPCR, multiplex qPCR or RT-qPCR, microarray analysis, SAGE, massARRAY technology, FISH, or combinations thereof. In some cases, mRNA expression levels are detected using RNA-seq.

In other cases, the expression level is a protein expression level. Protein expression levels can be determined using any suitable method (e.g., any of the methods disclosed herein). In some cases, protein expression levels are determined by IHC, immunofluorescence, mass spectrometry, flow cytometry, and western blotting, or combinations thereof.

Any suitable sample may be used. In some cases, the sample is a tissue sample, a cell sample, a whole blood sample, a plasma sample, a serum sample, or a combination thereof. In some cases, the tissue sample is a tumor tissue sample. In some cases, the tumor sample is a tumor tissue sample. In some cases, the tumor tissue sample comprises tumor cells, tumor infiltrating immune cells, stromal cells, NAT cells, or a combination thereof. In some cases, the tumor tissue sample is an FFPE sample, an archived sample, a fresh sample, or a frozen sample. In some cases, the tumor tissue sample is an FFPE sample.

The cancer may be any suitable type of cancer. In some cases, the cancer is lung cancer, kidney cancer, bladder cancer, breast cancer, colorectal cancer, ovarian cancer, pancreatic cancer, gastric cancer, esophageal cancer, mesothelioma, melanoma, head and neck cancer, thyroid cancer, sarcoma, prostate cancer, glioblastoma, cervical cancer, thymus cancer, leukemia, lymphoma, myeloma, mycosis fungoides, merkel cell carcinoma, or hematological malignancy. In some cases, the cancer is lung, kidney, bladder, or breast cancer. In some cases, the lung cancer is NSCLC. In some cases, the NSCLC is non-squamous NSCLC. In some cases, the NSCLC is squamous NSCLC.

In some cases, the benefit comprises a prolongation of OS in the individual, a prolongation of PFS in the individual, and/or an improvement in BCOR in the individual as compared to treatment without the PD-L1 axis binding antagonist. In some cases, the benefit includes an extension of the Overall Survival (OS) of the individual as compared to treatment without the PD-L1 axis binding antagonist.

(ii) Gene signature associated with plasma B cells

In some aspects, the B cell characteristic used in conjunction with the compositions and methods of the present disclosure is a plasma B cell characteristic. Any suitable plasma B cell characteristics may be used. For example, a plasma B cell characteristic can include one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14) genes listed in table 9.

Table 9: exemplary plasma B cell signature genes

MZB1
	DERL3
JSRP1
	TNFRSF17
SLAMF7
	IGHG2
IGHGP
	IGLV3-1
IGLV6-57
	IGHA2
IGKV4-1
	IGKV1-12
IGLC7
	IGLL5

For example, provided herein is a method of identifying an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) who is likely to benefit from treatment comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as astuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), the method comprising determining in a sample from the individual one or more of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or more of the genes are likely to be expressed at a high level in the individual, wherein the immune axis is expressed at a reference level that would benefit from treatment comprising expression of the individual at the reference level of the one or more of the individual.

In another example, provided herein is a method of selecting a therapy for an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising determining the expression level of one or more of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14) in a sample from the individual, wherein the individual is likely to comprise an anti-PD antibody, e.g., an anti-PD 1-PD antibody, e.g., an anti-PD-1, or an anti-PD antagonist.

In some cases, the method includes determining an expression level of one of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

In some cases, the method comprises determining an expression level of MZB 1.

In some cases, the method comprises determining an expression level of DERL 3.

In some cases, the method comprises determining an expression level of JSRP 1.

In some cases, the method includes determining the expression level of SLAMF 7.

In some cases, the method comprises determining the expression level of IGHG 2.

In some cases, the method comprises determining the expression level of IGHGP.

In some cases, the method comprises determining the expression level of IGLV 3-1.

In some cases, the method comprises determining the expression level of IGLV 6-57.

In some cases, the method comprises determining the expression level of IGHA 2.

In some cases, the method includes determining the expression level of IGKV 4-1.

In some cases, the method includes determining the expression level of IGKV 1-12.

In some cases, the method comprises determining the expression level of IGLC 7.

In some cases, the method includes determining the expression level of IGLL 5.

Any suitable immuno-score reference expression level can be used. In some cases, the immune score reference expression level is an immune score expression level of one or more genes in the reference population. In some cases, the reference population is a population of individuals with cancer. In some cases, the population of individuals includes a first subset of individuals who have received treatment with a PD-L1 axis binding antagonist and a second subset of individuals who have received treatment with a therapy that does not contain a PD-L1 axis binding antagonist. In some cases, the reference immune score expression level visibly differentiates each of the first subset of individuals from the second subset of individuals based on a significant difference between the individual's responsiveness to treatment with a PD-L1 axis binding antagonist and the individual's responsiveness to treatment with a therapy that does not contain a PD-L1 axis binding antagonist, above the reference immune score expression level, wherein the individual's responsiveness to treatment with a PD-L1 axis binding antagonist is significantly improved relative to the individual's responsiveness to treatment with a therapy that does not contain a PD-L1 axis binding antagonist. In some cases, the therapy without the PD-L1 axis binding antagonist comprises an anti-tumor agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, or a combination thereof. In some cases, the therapy without the PD-L1 axis binding antagonist comprises a chemotherapeutic agent. In some cases, the chemotherapeutic agent is docetaxel. In some cases, responsiveness to treatment comprises a prolongation of OS, a prolongation of PFS, or an increase in BCOR. In some cases, responsiveness to treatment includes an extension of Overall Survival (OS). In some cases, the reference immune score expression level is the median of the expression levels of each of the one or more genes in the reference population. In some cases, the median expression level is the median of the mean Z scores for the expression levels of each of the one or more genes in the reference population.

For example, provided herein is a method of identifying an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) who may benefit from treatment comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attrituzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), the method comprising determining in a sample from the individual one or more of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 11, 13, or 13) in which the individual is likely to benefit from treatment using a high level of expression of the PD-L1, wherein the gene comprises an immune-1 or more of the antagonist.

In another example, provided herein is a method of selecting a therapy for an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), renal cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising determining the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14) of the genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from the individual, wherein in the event that the immune score expression level of the one or more genes is greater than the reference immune score expression level of the one or more genes, identifying the individual as an individual likely to benefit from treatment comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attritumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), wherein the benefit comprises an increase in OS of the individual as compared to treatment without the PD-L1 axis binding antagonist.

In some cases, the genes include two or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

For example, provided herein is a method of identifying an individual having cancer who is likely to benefit from treatment comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as astuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), the method comprising determining the expression levels of two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14) of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from the individual, wherein the cases where the immune expression levels of the two or more genes are higher than a reference immune expression level of the two or more genes identify the individual as likely to benefit from treatment comprising a PD-L1 axis antagonist.

In another example, provided herein is a method of selecting a therapy for an individual having cancer, the method comprising determining the expression level of two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14) of the genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from the individual, wherein the individual is identified as an individual likely to benefit from treatment comprising a PD-L1 axis binding antagonist if the immune score expression level of the two or more genes is greater than a reference immune expression score level of the two or more genes.

In another example, provided herein is a method of identifying an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., NSCLC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) who may benefit from treatment comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attrituzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), the method comprising determining in a sample from the individual two or more of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, iguc 2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 genes) in which expression of the genes would benefit from treatment comprising a high level of immune binding to the individual, wherein the increased expression of the PD-L1 axis binding antagonist comprises a reference level of the PD, wherein the subject's would benefit from treatment comprising a high level of the increased expression of the PD-1 or a high level of the PD-1.

In yet another example, provided herein is a method of selecting a therapy for an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising determining the expression levels of two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14) of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from the individual, wherein in the event that the level of immune score expression of the two or more genes is greater than the reference level of immune score expression of the two or more genes, identifying the individual as an individual likely to benefit from treatment comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attritumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), wherein the benefit comprises a prolongation of OS in the individual as compared to treatment without the PD-L1 axis binding antagonist.

In some cases, the immune score expression level of the two or more genes in the sample is higher than the reference immune score expression level, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)). Any suitable PD-L1 axis binding antagonist can be administered, e.g., any PD-L1 axis binding antagonist provided herein (e.g., as described in section IV below).

Any combination of plasma B cell signature genes can be determined. For example, the combination may include two genes selected from MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5, e.g., any one of the combinations listed in Table 10. In another example, the combination may include three genes selected from MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5, e.g., any one of the combinations listed in Table 11. In another example, the combination may include four genes selected from MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5, e.g., any one of the combinations listed in table 12. In another example, the combination can include five genes selected from MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5, e.g., any one of the combinations listed in table 13. In another example, the combination may include six genes selected from MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5, e.g., any one of the combinations listed in table 14. In another example, the combination may include seven genes selected from MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5, e.g., any one of the combinations listed in Table 15.

Table 10: exemplary two-Gene combinations of plasma B cell signature genes

Table 11: exemplary three-Gene combinations of plasma B cell signature genes

MZB1, DERL3 and JSRP1
	MZB1, DERL3 and TNFRSF17
MZB1, DERL3 and SLAMF7
	MZB1, DERL3 and IGHG2
MZB1, DERL3 and IGHGP
	MZB1, DERL3 and IGLV3-1
MZB1, DERL3 and IGLV6-57
	MZB1, DERL3 and IGHA2
MZB1, DERL3 and IGKV4-1
	MZB1, DERL3 and IGKV1-12
MZB1, DERL3 and IGLC7
	MZB1, DERL3 and IGLL5
MZB1, JSRP1 and TNFRSF17
	MZB1, JSRP1 and SLAMF7
MZB1, JSRP1 and IGHG2
	MZB1, JSRP1 andIGHGP
MZB1, JSRP1 and IGLV3-1
	MZB1, JSRP1 and IGLV6-57
MZB1, JSRP1 and IGHA2
	MZB1, JSRP1 and IGKV4-1
MZB1, JSRP1 and IGKV1-12
	MZB1, JSRP1 and IGLC7
MZB1, JSRP1 and IGLL5
	MZB1, TNFRSF17 and SLAMF7
MZB1, TNFRSF17 and IGHG2
	MZB1, TNFRSF17 and IGHGP
MZB1, TNFRSF17 and IGLV3-1
	MZB1, TNFRSF17 and IGLV6-57
MZB1, TNFRSF17 and IGHA2
	MZB1, TNFRSF17 and IGKV4-1
MZB1, TNFRSF17 and IGKV1-12
	MZB1, TNFRSF17 and IGLC7
MZB1, TNFRSF17 and IGLL5

Table 12: exemplary four Gene combinations of plasma B cell signature genes

Table 13: exemplary five-Gene combinations of plasma B cell signature genes

MZB1, DERL3, JSRP1, TNFRSF17, and SLAMF7
	MZB1, DERL3, JSRP1, TNFRSF17, and IGHG2
MZB1, DERL3, JSRP1, TNFRSF17, and IGHGP
	MZB1, DERL3, JSRP1, TNFRSF17, and IGLV3-1
MZB1, DERL3, JSRP1, TNFRSF17, and IGLV6-57
	MZB1, DERL3, JSRP1, TNFRSF17, and IGHA2
MZB1, DERL3, JSRP1, TNFRSF17, and IGKV4-1
	MZB1, DERL3, JSRP1, TNFRSF17, and IGKV1-12
MZB1、DERL3、JSRP1, TNFRSF17 and IGLC7
	MZB1, DERL3, JSRP1, TNFRSF17, and IGLL5
MZB1, DERL3, JSRP1, SLAMF7, and IGHG2
	MZB1, DERL3, JSRP1, SLAMF7, and IGHGP
MZB1, DERL3, JSRP1, SLAMF7, and IGLV3-1
	MZB1, DERL3, JSRP1, SLAMF7 and IGLV6-57
MZB1, DERL3, JSRP1, SLAMF7 and IGHA2
	MZB1, DERL3, JSRP1, SLAMF7, and IGKV4-1
MZB1, DERL3, JSRP1, SLAMF7, and IGKV1-12
	MZB1, DERL3, JSRP1, SLAMF7, and IGLC7
MZB1, DERL3, JSRP1, SLAMF7, and IGLL5
	MZB1, DERL3, JSRP1, IGHG2 and IGHGP
MZB1, DERL3, JSRP1, IGHG2, and IGLV3-1
	MZB1, DERL3, JSRP1, IGHG2, and IGLV6-57
MZB1, DERL3, JSRP1, IGHG2, and IGHA2
	MZB1, DERL3, JSRP1, IGHG2 and IGKV4-1
MZB1, DERL3, JSRP1, IGHG2, and IGKV1-12
	MZB1, DERL3, JSRP1, IGHG2, and IGLC7
MZB1, DERL3, JSRP1, IGHG2, and IGLL5

Table 14: exemplary six-Gene combinations of plasma B cell signature genes

Table 15: exemplary seven Gene combinations of plasma B cell signature genes

In some cases, the genes include MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5.

The cancer may be any suitable type of cancer. In some cases, the cancer is lung cancer, kidney cancer, bladder cancer, breast cancer, colorectal cancer, ovarian cancer, pancreatic cancer, stomach cancer, esophageal cancer, mesothelioma, melanoma, head and neck cancer, thyroid cancer, sarcoma, prostate cancer, glioblastoma, cervical cancer, thymus cancer, leukemia, lymphoma, myeloma, mycosis fungoides, merkel cell carcinoma, or a hematological malignancy. In some cases, the cancer is lung, kidney, bladder, or breast cancer. In some cases, the lung cancer is NSCLC. In some cases, the NSCLC is non-squamous NSCLC. In some cases, the NSCLC is squamous NSCLC.

In some cases, the benefit comprises a prolongation of OS in the individual, a prolongation of PFS in the individual, and/or an improvement in BCOR in the individual as compared to treatment without the PD-L1 axis binding antagonist. In some cases, the benefit comprises an extension of Overall Survival (OS) of the individual as compared to treatment without the PD-L1 axis binding antagonist.

B. Tertiary Lymphatic Structure (TLS)

In some aspects, the methods provided herein can involve determining the presence of TLS in a sample (e.g., a tumor sample) from an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), renal cancer (e.g., RCC), and breast cancer (e.g., TNBC)).

For example, provided herein is a method of identifying an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) that is likely to benefit from treatment with a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), comprising determining the presence of a Tertiary Lymphoid Structure (TLS) in a sample (e.g., a tumor sample) from the individual, wherein the presence of the TLS in the sample identifies the individual as an individual that is likely to benefit from treatment with the PD-L1 axis binding antagonist.

In another example, provided herein is a method of selecting a therapy for an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising determining the presence of TLS in a sample (e.g., a tumor sample) from the individual, wherein the presence of TLS in the sample identifies the individual as an individual who is likely to benefit from treatment with a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attritumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In some cases, the presence of TLS is determined in a sample from the individual, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attritumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)). Any suitable PD-L1 axis binding antagonist can be administered, e.g., any PD-L1 axis binding antagonist provided herein (e.g., as described in section IV below).

Any suitable method can be used to determine the presence of TLS in a sample. For example, in some cases, the presence of TLS is determined by histological staining, IHC, immunofluorescence, or gene expression analysis.

Any suitable histological staining method may be used. For example, in some cases, histological staining includes hematoxylin and eosin (H & E) staining.

Any suitable IHC or immunofluorescence method may be used. In some cases, IHC or immunofluorescence includes detecting CD62L, L-selectin, CD40, or CD8, e.g., using antibodies (e.g., anti-CD 62L antibodies, anti-L-selectin antibodies, anti-CD 40 antibodies, and/or anti-CD 8 antibodies). In some cases, the MECA-79 antibody is used to detect CD62L or L-selectin.

In some cases, the gene expression analysis comprises determining the expression level of a TLS gene signature in the sample. For example, gene expression analysis can involve determining the expression level of any of the TLS features disclosed herein (see, e.g., section II, section C, below). In some cases, a TLS gene signature comprises one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13. In some cases, the gene comprises two or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13.

In some cases, the sample may include one TLS. In other cases, a sample can include more than one TLS, e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 50, 60, 70, 80, 90, 100, or more TLSs.

C.TLS feature

In some aspects, the methods provided herein can involve determining the expression level of one or more genes in the TLS signature. Any suitable TLS feature may be used. For example, the TLS signature can include one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) genes listed in table 16.

Table 16: exemplary TLS signature genes

CCL2
	CCL3
CCL4
	CCL5
CCL8
	CCL18
CCL19
	CCL21
CXCL9
	CXCL10
CXCL11
	CXCL13

For example, provided herein is a method of identifying an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) who may benefit from treatment comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attrituzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), the method comprising determining, in a sample from the individual, an expression level of one or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12), wherein an individual who may benefit from an immune expression level of the one or more genes is higher than an immune expression level of the reference axis binding antagonist, the individual would benefit from treatment comprising the PD-L1.

In another example, provided herein is a method of selecting a therapy for an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising determining an expression level of one or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) in a sample from the individual, wherein the individual is identified as likely to comprise a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 binding antagonist, e.g., an anti-PD-1 bead antibody, e.g., an anti-PD 1-1 monoclonal antibody)) for the treatment of the individual.

In some cases, the immune score expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) genes in the sample is greater than a reference immune score expression level of the one or more genes, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as alemtuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)). Any suitable PD-L1 axis binding antagonist can be administered, e.g., any PD-L1 axis binding antagonist provided herein (e.g., as described in section IV below).

Any suitable immuno-score reference expression level can be used. In some cases, the immune score reference expression level is an immune score expression level of one or more genes in the reference population. In some cases, the reference population is a population of individuals with cancer. In some cases, the population of individuals includes a first subset of individuals who have received treatment with a PD-L1 axis binding antagonist and a second subset of individuals who have received treatment with a therapy that does not contain a PD-L1 axis binding antagonist. In some cases, the reference immune score expression level visibly differentiates each of the first subset of individuals from the second subset of individuals based on a significant difference between the individual's responsiveness to treatment with a PD-L1 axis binding antagonist and the individual's responsiveness to treatment with a therapy that does not contain a PD-L1 axis binding antagonist, above the reference immune score expression level, wherein the individual's responsiveness to treatment with a PD-L1 axis binding antagonist is significantly improved relative to the individual's responsiveness to treatment with a therapy that does not contain a PD-L1 axis binding antagonist. In some cases, the therapy without the PD-L1 axis binding antagonist comprises an anti-tumor agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, or a combination thereof. In some cases, the therapy without the PD-L1 axis binding antagonist comprises a chemotherapeutic agent. In some cases, the chemotherapeutic agent is docetaxel. In some cases, responsiveness to treatment comprises prolongation of OS, prolongation of PFS, or an increase in BCOR. In some cases, responsiveness to treatment includes an extension of Overall Survival (OS). In some cases, the reference immune score expression level is the median of the expression levels of each of the one or more genes in the reference population. In some cases, the median expression level is the median of the mean Z scores for the expression levels of each of the one or more genes in the reference population.

In some cases, the gene comprises two or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13.

For example, provided herein is a method of identifying an individual having a cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) who is likely to benefit from treatment comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as astuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), comprising determining the expression levels of two or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) in a sample from the individual, wherein identifying the individual as having a PD-L1 axis binding antagonist is likely to benefit from treatment wherein the individual comprises identifying the reference expression level of the PD-L1 axis binding antagonist.

In another example, provided herein is a method of selecting a therapy for an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising determining an expression level of two or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) in a sample from the individual, wherein the individual is identified as likely to benefit from treatment of the individual comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attritrin bead antagonist) or an anti-PD 1 binding antibody, e.g., anti-PD 1 binding monoclonal antibody)).

In some cases, the immune score expression level of two or more genes in the sample is greater than the reference immune score expression level, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist. Any suitable PD-L1 axis binding antagonist can be administered, e.g., any PD-L1 axis binding antagonist provided herein (e.g., as described in section IV below).

In some cases, the reference immune score expression level is an immune score expression level of two or more genes in a reference population. In some cases, the reference population is a population of individuals with cancer. In some cases, the population of individuals includes a first subset of individuals who have received treatment with a PD-L1 axis binding antagonist and a second subset of individuals who have received treatment with a therapy that does not contain a PD-L1 axis binding antagonist. In some cases, the reference immune score expression level visibly differentiates each of the first subset of individuals from the second subset of individuals based on a significant difference between the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist and the responsiveness of the individual to treatment with the therapy without the PD-L1 axis binding antagonist above the reference expression level, wherein the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist is significantly improved relative to the responsiveness of the individual to treatment with the therapy without the PD-L1 axis binding antagonist. In some cases, the therapy without the PD-L1 axis binding antagonist comprises an anti-neoplastic agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, or a combination thereof. In some cases, the therapy without the PD-L1 axis binding antagonist comprises a chemotherapeutic agent. In some cases, the chemotherapeutic agent is docetaxel. In some cases, responsiveness to treatment comprises prolongation of OS, prolongation of PFS, or an increase in BCOR. In some cases, responsiveness to treatment includes an extension of Overall Survival (OS). In some cases, the reference immune score expression level is the median of the expression levels of each of the two or more genes in the reference population. In some cases, the median expression level is the median of the mean Z scores for the expression levels of each of the two or more genes in the reference population.

In some cases, the gene comprises three or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13. In some cases, the gene comprises four or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13. In some cases, the gene comprises five or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13. In some cases, the gene comprises six or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13. In some cases, the gene comprises seven or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13. In some cases, the gene comprises eight or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13. In some cases, the gene comprises nine or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13. In some cases, the gene comprises ten or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13. In some cases, the gene comprises eleven or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13.

Any combination of TLS signature genes can be determined, for example, as any combination of two genes selected from: CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13; any combination of three genes selected from: CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13; any combination of four genes selected from: CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13; any combination of five genes selected from: CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13; any combination of six genes selected from: CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13; any combination of seven genes selected from: CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13; any combination of eight genes selected from: CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13; any combination of nine genes selected from: CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13; any combination of ten genes selected from: CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13; any combination of eleven genes selected from: CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13; or any combination of eleven genes selected from: CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11 and CXCL13.

In some cases, the gene comprises CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13.

In other cases, the expression level is a protein expression level. Protein expression levels can be determined using any suitable method (e.g., any of the methods disclosed herein). In some cases, protein expression levels are determined by IHC, immunofluorescence, mass spectrometry, flow cytometry, and western blot, or a combination thereof.

B cell number and clonally expanded B cells

In some aspects, the methods provided herein can involve determining the presence and/or number of B cells in a sample from an individual. In some aspects, the methods provided herein can involve determining the presence and/or number of clonally expanded B cells in a sample from an individual.

For example, provided herein is a method of identifying an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) that is likely to benefit from treatment with a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), comprising determining the number of B cells in a sample (e.g., a tumor sample) from the individual, wherein the individual is identified as an individual that is likely to benefit from treatment with the PD-L1 axis binding antagonist if the number of B cells in the sample is greater than a reference number of B cells.

In another example, provided herein is a method of selecting a therapy for an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising determining the number of B cells in a sample (e.g., a tumor sample) from the individual, wherein the individual is identified as an individual who is likely to benefit from treatment with a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., altemazumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) where the number of B cells in the tumor sample is greater than a reference number of B cells.

In some cases, the number of B cells in the sample is greater than the reference number, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)). Any suitable PD-L1 axis binding antagonist can be administered, e.g., any PD-L1 axis binding antagonist provided herein (e.g., as described in section IV below).

The presence and/or quantity of any suitable type of B cell can be determined. For example, in some cases, B cells include CD79+ B cells, igG + B cells, and/or plasma cells.

In some embodiments, the presence and/or number of clonally expanded B cells may be determined.

For example, provided herein is a method of identifying an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) who may benefit from treatment comprising a PD-L1 axis binding antagonist (e.g., an anti-PD-L1 antibody, such as attentimab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), the method comprising determining whether the individual has clonally expanded B cells in a sample (e.g., a tumor sample) from the individual, wherein the clonally expanded B cells in the sample identify the individual as an individual who may benefit from treatment comprising a PD-L1 axis binding antagonist.

In another example, provided herein is a method of selecting a therapy for an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising determining whether the individual has clonally expanded B cells in a sample (e.g., a tumor sample) from the individual, wherein the clonally expanded B cells in the sample identify the individual as an individual likely to benefit from treatment with a PD-L1 axis binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attrituximab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In some cases, the sample (e.g., a tumor sample) comprises clonally expanded B cells, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attritumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

The clonally expanded B cells may be any type of B cells. For example, in some cases, the clonally expanded B cells are clonally expanded plasma cells.

Clonally expanded B cells may be detected by any suitable method. For example, in some cases, clonally expanded B cells are detected by measuring the diversity of B Cell Receptor (BCR) gene lineages in tumor samples. In some cases, where a Shannon Diversity Index (SDI) of a BCR gene lineage in a tumor sample from an individual is lower than a reference SDI, the individual is identified as an individual who is likely to benefit from treatment comprising a PD-L1 axis binding antagonist.

E.T Effector characteristics

Any of the methods described herein may further comprise determining the presence and/or expression level of one or more T effector characteristic genes. Any suitable T effector feature may be used. For example, the T effector characteristic may include one or more (e.g., 1, 2, 3, 4, 5, 6, 7, or 8) genes listed in table 17. In some cases, CD274 is further detected.

Table 17: exemplary T Effector signature genes

CD8A
	EOMES
GZMA
	TBX21
IFNG
	GZMB
CXCL9
	CXCL10

For example, provided herein is a method of identifying an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) that may benefit from treatment with a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attrituzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), the method comprising determining, in a sample from the individual, one or more of (i) the genes listed in table 1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) and (ii) the genes CD8A, EOMES, GZMA, TBX21, ifcl, cxb, CXCL and 10 (e.g., TNBC), 1, 2, 3, 4, 5, 6, 7, or 8), wherein the individual is identified as an individual likely to benefit from treatment with the PD-L1 axis binding antagonist in the event that the immune score expression level of (i) one or more of the genes listed in table 1 and (ii) one or more of the genes CD8A, EOMES, GZMA, TBX21, IFNG, GZMB, CXCL9, and CXCL10 is greater than the reference immune score expression level of one or more of the genes listed in table 1 and one or more of the genes CD8A, EOMES, GZMA, TBX21, IFNG, GZMB, CXCL9, and CXCL 10.

In another example, provided herein is a method of selecting a therapy for an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising determining in a sample from the individual (i) one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) of the genes listed in table 1 and (ii) one or more (e.g., 1, 2, 3, 4, 5, 6, 7, or 8), wherein the individual is identified as an individual who is likely to benefit from treatment with a PD-L1 axis binding antagonist (e.g., an anti-PD-L1 antibody, such as attritor mab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) if the immune score expression level of (i) one or more of the genes listed in table 1 and (ii) one or more of the genes CD8A, EOMES, GZMA, TBX21, ifcl 9, and CXCL10 is higher than the reference immune score expression level of one or more of the genes listed in table 1 and one or more of the genes CD8A, EOMES, GZMA, TBX21, IFNG, GZMB, CXCL9, and CXCL 10.

In some cases, the immune score expression level of one or more of (i) the genes listed in table 1 and (ii) one or more of the genes CD8A, EOMES, GZMA, TBX21, IFNG, GZMB, CXCL9, and CXCL10 in the sample is greater than the reference immune score expression level, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist. Any suitable PD-L1 axis binding antagonist can be administered, e.g., any PD-L1 axis binding antagonist provided herein (e.g., as described in section IV below).

In some cases, the reference immune score expression level is an immune score expression level of one or more of (i) one or more genes listed in table 1 and (ii) one or more of the genes CD8A, EOMES, GZMA, TBX21, IFNG, GZMB, CXCL9, and CXCL10 in the reference population. In some cases, the reference population is a population of individuals with cancer. In some cases, the population of individuals includes a first subset of individuals who have received treatment with a PD-L1 axis binding antagonist and a second subset of individuals who have received treatment with a therapy that does not contain a PD-L1 axis binding antagonist. In some cases, the reference immune score expression level clearly distinguishes each of the first subset of individuals from the second subset of individuals based on a significant difference between the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist and the responsiveness of the individual to treatment with the therapy that does not contain the PD-L1 axis binding antagonist above the reference expression level, where the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist is significantly improved relative to the responsiveness of the individual to treatment with the therapy that does not contain the PD-L1 axis binding antagonist. In some cases, the therapy without the PD-L1 axis binding antagonist comprises an anti-neoplastic agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, or a combination thereof. In some cases, the therapy without the PD-L1 axis binding antagonist comprises a chemotherapeutic agent. In some cases, the chemotherapeutic agent is docetaxel. In some cases, responsiveness to treatment comprises prolongation of OS, prolongation of PFS, or an increase in BCOR. In some cases, responsiveness to treatment includes an extension of Overall Survival (OS). In some cases, the reference immune score expression level is the median of the expression levels of each of the two or more genes in the reference population. In some cases, the median expression level is the median of the mean Z scores for the expression levels of each of (i) the one or more genes listed in table 1 and (ii) the genes CD8A, EOMES, GZMA, TBX21, IFNG, GZMB, CXCL9, and CXCL10 in the reference population.

In another example, provided herein is a method of identifying an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., NSCLC), kidney cancer (e.g., rcuc), and breast cancer (e.g., TNBC)) that may benefit from treatment with a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), the method comprising determining in a sample from the individual (i) one or more of B-cell signature genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) and (ii) the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, or 8) of the T effector feature genes CD8A, EOMES, GZMA, TBX21, IFNG, GZMB, CXCL9, and CXCL10, wherein the individual is identified as an individual likely to benefit from treatment with the PD-L1 axis binding antagonist if (i) the one or more B cell feature genes and (ii) the one or more T effector feature genes are greater than the reference immune score expression level of the one or more B cell feature genes and the one or more T effector feature genes.

In yet another example, provided herein is a method of selecting a therapy for an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising determining in a sample from the individual one or more of (i) B cell signature genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) and (ii) T effector signature genes CD8A, EOMES, GZMA, TBX21, IFNG, gzm B, CXCL9, and CXCL10 (e.g., 1, 2, 3, 4, 5, 6, 7, or 8 antagonists) wherein the expression of the one or more of the B cell signature genes (e.g., PD) is likely to be expressed at a level that is higher than the level of the anti-PD 1-effector gene, e.g., when the anti-PD gene is expressed by the individual, and the anti-PD receptor(s) and the anti-PD) signature genes are expressed at a level that is high, e.g., when the anti-PD 1-PD gene is likely to the anti-PD gene.

In some cases, the immune score expression level of (i) the one or more B cell signature genes and (ii) the one or more T effector signature genes in the sample is greater than a reference immune score expression level, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist. Any suitable PD-L1 axis binding antagonist can be administered, e.g., any PD-L1 axis binding antagonist provided herein (e.g., as described in section IV below).

In some cases, the reference immune score expression level is an immune score expression level of (i) the one or more B cell signature genes and (ii) the one or more T effector signature genes in the reference population. In some cases, the reference population is a population of individuals with cancer. In some cases, the population of individuals includes a first subset of individuals who have received treatment with a PD-L1 axis binding antagonist and a second subset of individuals who have received treatment with a therapy that does not contain a PD-L1 axis binding antagonist. In some cases, the reference immune score expression level clearly distinguishes each of the first subset of individuals from the second subset of individuals based on a significant difference between the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist and the responsiveness of the individual to treatment with the therapy that does not contain the PD-L1 axis binding antagonist above the reference expression level, where the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist is significantly improved relative to the responsiveness of the individual to treatment with the therapy that does not contain the PD-L1 axis binding antagonist. In some cases, the therapy without the PD-L1 axis binding antagonist comprises an anti-neoplastic agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, or a combination thereof. In some cases, the therapy without the PD-L1 axis binding antagonist comprises a chemotherapeutic agent. In some cases, the chemotherapeutic agent is docetaxel. In some cases, responsiveness to treatment comprises a prolongation of OS, a prolongation of PFS, or an increase in BCOR. In some cases, responsiveness to treatment includes an extension of Overall Survival (OS). In some cases, the reference immune score expression level is the median of the expression levels of each of the two or more genes in the reference population. In some cases, the median expression level is the median of the mean Z-scores of the expression levels of each of (i) the one or more B cell signature genes and (ii) the one or more T effector signature genes in the reference population.

In another example, provided herein is a method of identifying an individual having a cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) who is likely to benefit from treatment comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as alemtuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), comprising determining in a sample from the individual the level of (i) one or more of the TLS signature genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, cl9, CXCL10, CXCL11, and CXCL13 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) and (ii) the T effector signature gene antagonist (e.g., CD8A, eoa, mes, 10, 11, or 12) and (ii) in which the individual is likely to benefit from a treatment comprising one or more of PD-L1 axis binding of the PD-L1, 2, gmc, or TNBC) and a high level of the signature gene.

In yet another example, provided herein is a method of selecting a therapy for an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising determining in a sample from the individual one or more of (i) the TLS signature genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) and (ii) the T effector signature genes CD8A, EOMES, GZMA, TBX21, IFNG, GZMB, CXCL9, and CXCL10 (e.g., 1, 2, 3, 4, 5, 6, 7, or 12) and (ii) wherein the level of one or more of the anti-TLS signature genes (e.g., binding to the anti-PD antagonist) is expressed at a level that would benefit from treatment with the anti-PD 1 or more of the anti-PD 1-PD antagonist, e.g., PD, e.g., anti-PD, and anti-PD 1, e.g., anti-PD, and a monoclonal antibody.

In some cases, the immune score expression level of (i) the one or more TLS signature genes and (ii) the one or more T effector signature genes in the sample is greater than a reference immune score expression level, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist. Any suitable PD-L1 axis binding antagonist can be administered, e.g., any PD-L1 axis binding antagonist provided herein (e.g., as described in section IV below).

In some cases, the reference immune score expression level is the immune score expression level of (i) the one or more TLS signature genes and (ii) the one or more T effector signature genes in the reference population. In some cases, the reference population is a population of individuals with cancer. In some cases, the population of individuals includes a first subset of individuals who have received treatment with a PD-L1 axis binding antagonist and a second subset of individuals who have received treatment with a therapy that does not contain a PD-L1 axis binding antagonist. In some cases, the reference immune score expression level clearly distinguishes each of the first subset of individuals from the second subset of individuals based on a significant difference between the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist and the responsiveness of the individual to treatment with the therapy that does not contain the PD-L1 axis binding antagonist above the reference expression level, where the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist is significantly improved relative to the responsiveness of the individual to treatment with the therapy that does not contain the PD-L1 axis binding antagonist. In some cases, the therapy without the PD-L1 axis binding antagonist comprises an anti-neoplastic agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, or a combination thereof. In some cases, the therapy without the PD-L1 axis binding antagonist comprises a chemotherapeutic agent. In some cases, the chemotherapeutic agent is docetaxel. In some cases, responsiveness to treatment comprises prolongation of OS, prolongation of PFS, or an increase in BCOR. In some cases, responsiveness to treatment includes an extension of Overall Survival (OS). In some cases, the reference immune score expression level is the median of the expression levels of each of the two or more genes in the reference population. In some cases, the median expression level is the median of the mean Z scores for the expression levels of each of (i) the one or more TLS signature genes and (ii) the one or more T effector signature genes in the reference population.

In some examples of any of the foregoing methods, the presence and/or level of expression of CD274 is further determined.

In any of the methods described herein, the method can further comprise determining the expression level of CD79A, CD274, and the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, or 8) of the T effector feature genes CD8A, EOMES, GZMA, TBX21, IFNG, GZMB, CXCL9, and CXCL 10.

F. Determination of expression levels

(i) Detection method

The level of immune score expression of a gene described herein (e.g., one or more genes listed in any of tables 1-17) can be based on the level of nucleic acid expression, and preferably on the level of mRNA expression. The presence and/or expression level/amount of a gene described herein can be determined qualitatively and/or quantitatively based on any suitable standard known in the art, including but not limited to DNA, mRNA, cDNA, protein fragment, and/or gene copy number.

In some cases, the nucleic acid expression level of a gene described herein (e.g., one or more genes listed in any one of tables 1-17) can be measured by Polymerase Chain Reaction (PCR) -based assays (e.g., quantitative PCR, real-time PCR, quantitative real-time PCR (qRT-PCR), reverse transcriptase PCR (RT-PCR), and reverse transcriptase quantitative PCR (RT-qPCR)). Platforms for performing quantitative PCR assays include (e.g., BIOMARK) ^TM HD system). Other amplification-based methods include, for example, transcription-mediated amplification (TMA), strand Displacement Amplification (SDA), nucleic acid sequence-based amplification (NASBA), and signal amplification methods such as bDNA.

In some cases, the nucleic acid expression levels of the genes described herein (e.g., one or more of the genes listed in any of tables 1-17) can also be determined by sequencing-based techniques such as RNA-seq, serial Analysis of Gene Expression (SAGE), high-throughput sequencing techniques (e.g., massively parallel sequencing), and Sequenom

The technique takes a measurement. Nucleic acid expression levels (e.g., expression levels of one or more of the genes listed in any of tables 1-17) can also be measured by, for example, nanoString nCounter and high coverage expression profiling (HiCEP). Other Protocols for assessing the status of genes and gene products can be found, for example, in Ausubel et al, 1995, current Protocols In Molecular Biology, unit 2 (Northern blot), 4 (Southern blot), 15 (immunoblot) and 18 (PCR analysis).

Other methods for detecting the nucleic acid square of a gene described herein (e.g., one or more genes listed in any of tables 1-17) include protocols for examining or detecting mRNA (such as a target mRNA) in a tissue or cell sample by microarray technology. Test and control mRNA samples from the test and control tissue samples were reverse transcribed and labeled using a nucleic acid microarray to generate cDNA probes. The probes are then hybridized to an array of nucleic acids immobilized on a solid support. The array is configured so that the order and location of each component of the array is known. Hybridization of a labeled probe to a particular array element indicates that the sample from which the probe was derived expresses the gene.

The primers and probes may be labeled with a detectable marker, such as a radioisotope, a fluorescent compound, a bioluminescent compound, a chemiluminescent compound, a metal chelator or an enzyme. Such probes and primers may be used to detect the presence of a gene expressed in a sample, such as one or more of the genes listed in any one of tables 1 to 17. As will be understood by those skilled in the art, many different primers and probes can be prepared based on the sequences provided herein (or their adjacent sequences in the case of genomic DNA) and used effectively to amplify, clone and/or determine the presence of and/or the expression level of the genes described herein.

Other methods for detecting the level of nucleic acid expression of a gene described herein (e.g., one or more genes listed in any one of tables 1 to 17) include electrophoresis, northern blot and Southern blot analysis, in situ hybridization (e.g., single or multiplex nucleic acid in situ hybridization), rnase protection assays, and microarrays (e.g., illumina BEADARRAY) ^TM Technology); bead Arrays (BADGE)) for detecting gene expression.

In some cases, the level of immune score expression of a gene described herein (e.g., one or more genes listed in any one of tables 1-17) can be analyzed by a variety of methods, including but not limited to RNA-seq, PCR, RT-qPCR, multiplex RT-qPCR,

Gene expression determination, microarray analysis, serial Analysis of Gene Expression (SAGE), northern blottingAnalysis, massARRAY, ISH, and whole genome sequencing, or a combination thereof.

In further instances, the level of immune score expression of a gene described herein (e.g., one or more genes listed in any one of tables 1-17) in a sample can be detected using a method selected from the group consisting of: RNA-seq, RT-qPCR, multiplex RT-qPCR, microarray analysis, SAGE, massARRAY technology, FACS, western blot analysis, ELISA, immunoprecipitation, immunohistochemistry, immunofluorescence, radioimmunoassay, dot hybridization, immunodetection methods, HPLC, surface plasmon resonance, spectroscopy, mass spectrometry, HPLC, and ISH, or combinations thereof.

(ii)RT-qPCR

In some cases, the level of nucleic acid expression of a gene described herein (e.g., one or more genes listed in any one of tables 1 to 17) can be detected using reverse transcription quantitative polymerase chain reaction (RT-qPCR). The RT-qPCR technique is a form of PCR in which the nucleic acid to be amplified is RNA that is first reverse transcribed into cDNA, and the amount of PCR product is measured in each step of the PCR reaction. Since RNA cannot be used as a template for PCR, the first step in gene expression profiling by PCR is to reverse transcribe the RNA template into cDNA, which is then amplified in a PCR reaction. For example, the reverse transcriptase can include avian myeloblastosis virus reverse transcriptase (AMY-RT) or Moloney murine leukemia virus reverse transcriptase (MMLV-RT). Depending on the circumstances and the goal of expression profiling, the reverse transcription step is usually primed using specific primers, random hexamers or oligo-dT primers. For example, GENEAMP can be used ^TM RNA PCR kit (Perkin Elmer, calif, USA), reverse transcription of the extracted RNA was performed according to the manufacturer's instructions. The resulting cDNA can then be used as a template in subsequent PCR reactions.

One variation of the PCR technique is real-time quantitative PCR (qRT-PCR), which is performed by means of fluorescent probes (i.e.with double labelling)

Probe) to measure PCR product accumulation. Quantitative real-time polymerase chain reaction technique refers to PCRA format wherein the amount of PCR product is measured in each step of the PCR reaction. In the examples including Cronin et al, am.J.Pathol.164 (l): 35-42 (2004); and Ma et al, cancer Cell 5, 607-616 (2004). Real-time fluorescent quantitative PCR is compatible with quantitative competitive PCR (in which normalization is performed using internal competitor genes for each target sequence) and/or with quantitative comparative PCR (which performs PCR using normalization genes or housekeeping genes contained in a sample). For further details, see, e.g., held et al, genome Research 6, 986-994 (1996).

The steps of a representative protocol for analyzing gene expression using fixed, paraffin-embedded tissues as a source of RNA, including mRNA isolation, purification, primer extension and amplification, are provided in various published journal articles (e.g., godfrey et al, malec. Diagnostics 2. Briefly, one representative method begins by cutting a portion (e.g., a 10 microgram portion) of a paraffin-embedded tumor tissue sample. RNA is then extracted, and proteins and DNA are removed. After analysis of the RNA concentration, if necessary, RNA repair and/or amplification steps may be included, and the RNA is reverse transcribed using a gene-specific promoter, followed by PCR.

The level of nucleic acid expression determined by amplification-based methods (e.g., RT-qPCR) can be expressed as the cycle threshold (Ct). From this value, the normalized expression level of each gene can be determined using the Δ Ct (dCt) method, as follows: ct (control/reference gene) -Ct (target/target gene) = dCt (target/target gene). Those skilled in the art will appreciate that the dCt value obtained may be a negative dCt value, or may be a positive dCt value. As defined herein, a higher dCt value indicates a higher expression level of the gene of interest relative to a control gene. Conversely, a lower dCt value indicates a lower expression level of the gene of interest relative to the control gene. Where the expression levels of multiple genes have been determined, the expression level of each gene (e.g., expressed as a dCt value) can then be used to determine a single value (e.g., an immune score expression level) that represents the collective or composite expression level of the multiple genes. The immune score expression level can be the mean or median of the dCt values measured for each target/gene of interest. Thus, in some cases, an immune score expression level described herein can be the mean or median of dCt values measured for one or more of the genes listed in any of tables 1 to 17. As defined herein, a higher mean dCt or median dCt value indicates a higher overall expression level of the plurality of target genes relative to the control gene(s). A lower mean dCt or dCt median value indicates a lower overall level of expression of the plurality of target genes relative to the control gene(s). The immune score expression level can then be compared to a reference immune score expression level as further defined herein, as described herein.

In some embodiments, the expression level of a nucleic acid described herein can be determined using a method comprising: (a) Obtaining or providing a sample from an individual, wherein the sample comprises a tumor tissue sample (e.g., a formalin-fixed, paraffin-embedded NSCLC, UC, RCC, or TNBC tumor tissue sample); (b) isolating mRNA from the sample; (c) Reverse transcription of mRNA into cDNA (e.g., for one or more genes listed in any one of tables 1 to 17); (d) Amplifying cDNA using PCR (e.g., for one or more genes listed in any one of tables 1 to 17); and (e) quantifying the nucleic acid expression level (e.g., for one or more genes listed in any one of tables 1 to 17).

Depending on the primers or probes used, one or more (e.g., one or more genes listed in any of tables 1-17) can be detected in a single assay. Further, the assay can be performed across one or more test tubes (e.g., one, two, three, four, five, or six or more test tubes).

In some cases, the method further comprises: (f) The nucleic acid expression level of one or more genes (e.g., one or more genes listed in any of tables 1-17) in the sample is normalized to the expression level of one or more reference genes (e.g., one, two, three, four, five, six, seven, eight, nine or more reference genes, e.g., housekeeping genes). For example, RT-qPCR can be used to analyze the immune score expression levels of genes described herein (e.g., one or more genes listed in any one of tables 1-17) to generate immune score expression levels that reflect normalized mean dCT values for the analyzed genes.

(iii)RNA-seq

In some cases, the level of nucleic acid expression of a gene described herein (e.g., one or more genes listed in any one of tables 1-17) can be detected using RNA-seq. RNA-seq, also known as Whole Transcriptome Shotgun Sequencing (WTSS), refers to sequencing and/or quantifying cDNA using high-throughput sequencing techniques in order to obtain information about the RNA content of a sample. Publications describing RNA-Seq include: wang et al, RNA-Seq: a reliable tool for transcriptors, nature Reviews Genetics 10 (1): 57-63 (1, 2009); ryan et al, bioTechniques 45 (1): 81-94 (2008); and Maher et al, transgenic sequences to detection genes in cander, nature 458 (7234): 97-101 (1 month 2009).

(iv) Sample (I)

The samples described herein can be taken, for example, from individuals suspected of having or diagnosed with cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) and thus may be in need of treatment, or from healthy individuals not suspected of having cancer or determined to not have cancer but a family history of cancer. To assess gene expression, samples, such as those comprising cells or proteins or nucleic acids produced by such cells, can be used in the methods of the invention. The expression level of a gene can be determined by assessing the amount (e.g., absolute amount or concentration) of a marker in a sample (e.g., a tissue sample, e.g., a tumor tissue sample, such as a biopsy sample). In addition, the gene level in the bodily fluid or excreta containing detectable gene levels can be assessed. Body fluids or secretions useful as samples in the present invention include, for example, blood, urine, saliva, stool, pleural fluid, lymph, sputum, ascites, prostatic fluid, cerebrospinal fluid (CSF) or any other body fluid or derivative thereof. The term "blood" is intended to include whole blood, plasma, serum or any derivative of blood. In cases where invasive sampling methods are inappropriate or inconvenient, it may sometimes be preferable to assess genes in such body fluids or excreta. In other embodiments, tumor tissue samples are preferred.

The sample can be a frozen sample, a fresh sample, a fixed (e.g., formalin fixed) sample, a centrifuged and/or embedded (e.g., paraffin embedded) sample, and the like. Prior to assessing the amount of one or more markers in a sample, the cell sample may be subjected to various well-known post-collection preparation and storage techniques (e.g., nucleic acid and/or protein extraction, immobilization, storage, freezing, ultrafiltration, concentration, evaporation, centrifugation, etc.). Likewise, the biopsy sample may also be processed by post-collection preparation and storage techniques (e.g., fixation, such as formalin fixation).

In some embodiments, the sample is a blood sample. In another instance, the sample is used in a diagnostic assay such as the diagnostic assay or diagnostic method of the invention. In some cases, the sample is obtained from a primary or metastatic tumor. Tissue biopsies are commonly used to obtain representative tumor tissue masses. Alternatively, tumor cells may be obtained indirectly in the form of tissues or body fluids known or believed to contain the tumor cells of interest. For example, samples of lung cancer lesions may be obtained by resection, bronchoscopy, fine needle puncture, bronchial brushing, or sputum, pleural fluid, or blood. Genes or gene products can be detected from cancer or tumor tissue or other body samples (e.g., urine, sputum, serum, or plasma). The same techniques discussed above for detecting a target gene or gene product in a cancerous sample can be applied to other body samples. Cancer cells may be shed from cancerous lesions and appear in such body samples. By screening such body samples, a simple early diagnosis of these cancers can be made. In addition, by testing for target genes or gene products in such body samples, the progress of the treatment can be more easily monitored.

In some cases, the sample from the individual is a tissue sample, a whole blood sample, a plasma sample, a serum sample, or a combination thereof. In some cases, the sample is a tissue sample. In some cases, the sample is a tumor tissue sample. In some cases, the sample is obtained prior to treatment with a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)). In some cases, the tissue sample is a Formalin Fixed and Paraffin Embedded (FFPE) sample, an archived sample, a fresh sample, or a frozen sample.

In some cases, the sample from the individual is a tissue sample. In some cases, the tissue sample is a tumor tissue sample (e.g., biopsy). In some cases, the tumor tissue sample comprises tumor cells, tumor infiltrating immune cells, stromal cells, or a combination thereof. In some cases, the tissue sample is lung tissue. In some cases, the tissue sample is bladder tissue. In some cases, the tissue sample is kidney tissue. In some cases, the tissue sample is breast tissue. In some cases, the tissue sample is skin tissue. In some cases, the tissue sample is pancreatic tissue. In some cases, the tissue sample is stomach tissue. In some cases, the tissue sample is esophageal tissue. In some cases, the tissue sample is mesothelial tissue. In some cases, the tissue sample is thyroid tissue. In some cases, the tissue sample is colorectal tissue. In some cases, the tissue sample is head or neck tissue. In some cases, the tissue sample is osteosarcoma tissue. In some cases, the tissue sample is prostate tissue. In some cases, the tissue sample is ovarian tissue, HCC (liver), blood cells, lymph nodes, or bone/bone marrow.

In some cases, the tumor tissue sample is extracted from a malignant tumor (i.e., cancer). In some cases, the cancer is a solid tumor or a non-solid tumor or a soft tissue tumor. Examples of soft tissue tumors include leukemia (e.g., chronic myelogenous leukemia, acute myelogenous leukemia, adult acute lymphocytic leukemia, acute myelogenous leukemia, mature B-cell acute lymphocytic leukemia, chronic lymphocytic leukemia, multi-lymphocytic leukemia, or hairy cell leukemia) or lymphoma (e.g., non-hodgkin's lymphoma, cutaneous T-cell lymphoma, or hodgkin's disease). Solid tumors include cancers of any body tissue other than the blood, bone marrow, or lymphatic system. Solid tumors can be further divided into epithelial cell-derived and non-epithelial cell-derived. Examples of epithelial solid tumors include gastrointestinal cancer, colon cancer, colorectal cancer (e.g., basal-like colorectal cancer), breast cancer, prostate cancer, lung cancer, kidney cancer, liver cancer, pancreatic cancer, ovarian cancer (e.g., endometrioid ovarian cancer), head and neck cancer, oral cancer, stomach cancer, duodenal cancer, small intestine cancer, large intestine cancer, rectal cancer, gallbladder cancer, lip cancer, nasopharyngeal cancer, skin cancer, uterine cancer, cancer of the male reproductive organ, cancer of the urinary organ (e.g., urothelial cancer, dysplastic urothelial cancer, transitional cell cancer), bladder cancer, and skin cancer. Solid tumors of non-epithelial origin include sarcomas, brain tumors and bone tumors. In some cases, the cancer is NSCLC. In some cases, the cancer is second or third line locally advanced or metastatic non-small cell lung cancer. In some cases, the cancer is adenocarcinoma. In some cases, the cancer is squamous cell carcinoma.

(v) RNA extraction

mRNA can be isolated from a target sample prior to detecting the level of nucleic acid. In some cases, the mRNA is total RNA isolated from a tumor or tumor cell line or a normal tissue or cell line. RNA can be isolated from various tumor tissues (including breast, lung, colon, prostate, brain, liver, kidney, pancreas, stomach, gall bladder, spleen, thymus, testis, ovary, uterus, etc.), corresponding normal tissues or tumor cell lines. If the source of the mRNA is a primary tumor, the mRNA can be extracted from frozen or archived paraffin-embedded and fixed (e.g., formalin-fixed) tissue samples. General methods for extracting mRNA are well known in the art and are disclosed in standard textbooks of molecular biology, including: ausubel et al, current Protocols of Molecular Biology, john Wiley and Sons (1997). Methods for extracting RNA from paraffin-embedded tissues are disclosed, for example, in the following documents: rupp and Locker, lab invest.56: A67 (1987); and De Andres et al, bio technologies 18 (1995). In particular, commercial manufacturers (e.g., qiagen) purification kits, buffer sets, and proteases can be used RNA isolation was performed according to the manufacturer's instructions. For example, total RNA can be extracted from cells in culture using Qiagen RNeasy mini-columns. Other commercially available RNA isolation kits include

Complete set of DNA and RNA purification kits (

Madison, wis.) and paraffin block RNA isolation kit (Ambion, inc.). For example, RNA Stat-60 (TelTest) can be used to isolate total RNA from tissue samples. For example, cesium chloride density gradient centrifugation can also be used to isolate RNA prepared from tumor tissue samples.

(vi) Immune score expression level

The immune score expression level may reflect the expression level of one or more genes described herein (e.g., one or more genes listed in any one of tables 1-17). In some cases, to determine the immune score expression level, the detected expression level of each gene is normalized using any of the standard normalization methods known in the art. One skilled in the art will appreciate that the normalization method used may depend on the gene expression method used (e.g., in the case of the RT-qPCR method, normalization may be performed using one or more housekeeping genes, but in the case of the RNA-seq method, the whole genome or substantially the whole genome may be used as a normalization baseline). For example, the detected expression levels of each gene tested can be normalized for differences in the amount of one or more genes tested, variability in the quality of the sample used, and/or variability between test runs.

In some cases, normalization can be achieved by detecting the expression of a particular normalization gene or genes (including one or more reference genes, such as housekeeping genes). For example, in some cases, the expression levels of nucleic acids detected using the methods described herein (e.g., for one or more genes listed in any one of tables 1-17) can be normalized to the expression levels of one or more reference genes (e.g., one, two, three, four, five, six, seven, eight, nine, or more reference genes, such as housekeeping genes). Alternatively, normalization can be based on the average signal or median of the signals for all genes tested. The normalized amount of tumor mRNA measured from the subject can be compared to the amount found at a reference immune score expression level on a gene-by-gene basis. The presence and/or expression level/amount measured in a particular subject sample to be analyzed will fall within a certain percentile of this range, which can be determined by methods well known in the art.

In some cases, the level of expression detected for each gene tested is not normalized for determining the level of immune score expression.

The immune score expression level may reflect a single gene or a collective or composite expression level of multiple genes described herein (e.g., for one or more genes listed in any one of tables 1-17). Any statistical method known in the art can be used to determine the level of expression of an immune score.

For example, the immune score expression level may reflect a median expression level, an average expression level, or a numerical value reflecting a collective Z-score expression level for a combination of genes determined (e.g., for one or more genes listed in any of tables 1-17).

In some cases, the immune score expression level reflects a median normalized expression level, an average normalized expression level, or a numerical value reflecting a collective Z-score normalized expression level for the combination of genes determined (e.g., for one or more genes listed in any one of tables 1-17).

For example, the immune score expression level may reflect the average (mean) of the expression levels of each gene in a combination of two or more genes listed in any one of tables 1 to 17. In some cases, the immune score expression level reflects an average (mean) of the normalized expression levels of each gene in a combination of two or more genes listed in any one of tables 1 to 17 (e.g., normalized to a reference gene, such as a housekeeping gene). In some cases, the immune score expression level reflects the median of the expression level of each gene in a combination of two or more genes listed in any one of tables 1 to 17. In some cases, the immune score expression level reflects the median value of the normalized expression level of each gene in a combination of two or more genes listed in any one of tables 1 to 17 (e.g., normalized to a reference gene, such as a housekeeping gene). In some cases, the immune score expression level reflects the Z score of each gene in a combination of two or more genes listed in any one of tables 1 to 17.

(vii) Reference immune score expression level

The reference immune score expression level can be a value obtained by analyzing any of the reference populations described herein. In some cases, the reference immune score expression level can be a "cut-off value selected based on the reference immune score expression level that divides the reference population into different subsets, e.g., subsets that have a significant difference (e.g., a statistically significant difference) in therapeutic response to PD-L1 axis binding antagonist therapy and non-PD-L1 axis binding antagonist therapy. In such cases, relative therapeutic response may be assessed based on progression-free survival (PFS) or Overall Survival (OS), e.g., expressed as a risk ratio (HR) (e.g., progression-free survival HR (PFS HR) or overall survival HR (OS HR)).

In certain instances, the reference immune score expression level is an immune score expression level of one or more genes listed in any one of tables 1 to 17 in the reference population that, based on a significant difference between the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist and the responsiveness of the individual to treatment with the non-PD-L1 axis binding antagonist that is greater than the reference immune score expression level (i.e., greater than a cutoff value), significantly (e.g., statistically) distinguishes a first subset of individuals in the reference population that have received treatment with the PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attlizumab) or a PD-1 binding antagonist (e.g., anti-PD-1 antibody)) from a second subset of individuals in the same reference population that have received treatment with the non-PD-L1 axis binding antagonist without the PD-L1 axis binding antagonist, wherein the individual's responsiveness to treatment with the PD-L1 axis binding antagonist is significantly improved relative to the individual's treatment with the non-PD-L1 axis binding antagonist.

In some cases, the reference immune score expression level is an immune score expression level of one or more genes listed in any one of tables 1 to 17 in the reference population that substantially best distinguishes a first subset of individuals in the reference population that have been treated with a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as alemtuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) from a second subset of individuals in the same reference population that have been treated with a non-PD-L1 axis binding antagonist that does not contain a PD-L1 axis binding antagonist, based on a substantially maximal difference between the individual's responsiveness to treatment with the PD-L1 axis binding antagonist and the individual's responsiveness to treatment with the non-PD-L1 axis binding antagonist (i.e., above a threshold), wherein the individual's responsiveness to treatment with the PD-L1 axis binding antagonist is statistically significantly improved (e.g., has a statistically significant response) to treatment with the non-PD-L1 axis binding antagonist.

In certain particular instances, the reference immune score expression level is an immune score expression level of one or more genes listed in any one of tables 1 to 17 in the reference population that is based on a maximum difference between the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist therapy and the responsiveness of the individual to treatment with the non-PD-L1 axis binding antagonist therapy that is greater than the reference immune score expression level (i.e., greater than a cutoff value), that best distinguishes a first subset of individuals in the reference population that have received treatment with the PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., alemtuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) from a second subset of individuals in the same reference population that have received treatment with the non-PD-L1 axis binding antagonist without the PD-L1 axis binding antagonist, wherein the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist therapy is significantly (e.g., statistically significantly improved) relative to treatment with the non-PD-L1 axis binding antagonist.

In certain instances, the reference immune score expression level is an immune score expression level of one or more genes listed in any one of tables 1 to 17 in the reference population that, based on a significant difference below the reference immune score expression level (i.e., below a threshold) between the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist and the responsiveness of the individual to treatment with the non-PD-L1 axis binding antagonist, visibly (e.g., statistically significant) distinguishes a first subset of individuals in the reference population that have been treated with the PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as alemtuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) from a second subset of individuals in the same reference population that have been treated with the non-PD-L1 axis binding antagonist without PD-L1 axis binding, wherein the responsiveness of the individual to treatment with the non-PD-L1 axis binding antagonist is statistically significantly improved (e.g., statistically significantly improved) relative to the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist.

In some cases, the reference immune score expression level is an immune score expression level of one or more genes listed in any one of tables 1 to 17 in the reference population that substantially optimally distinguishes a first subset of individuals in the reference population that have received treatment with a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., alemtuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) from a second subset of individuals in the same reference population that have received treatment with a non-PD-L1 axis binding antagonist that does not contain a PD-L1 axis binding antagonist, based on a substantially maximal difference between the individual's responsiveness to treatment with the PD-L1 axis binding antagonist and the individual's responsiveness to treatment with a non-PD-L1 axis binding antagonist (e.g., below a reference immune score expression level (i.e., below a cut-off value), wherein the individual's responsiveness to treatment with the PD-L1 axis binding antagonist is statistically significantly improved (e.g., has a statistically significant response to treatment with the PD-L1 axis binding antagonist).

In certain particular instances, the reference immune score expression level is an immune score expression level of one or more genes listed in any one of tables 1 to 17 in the reference population that is based on a maximum difference below the reference immune score expression level (i.e., below a threshold) between the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist therapy and the responsiveness of the individual to treatment with the non-PD-L1 axis binding antagonist therapy that best distinguishes a first subset of individuals in the reference population that have received treatment with the PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., alemtuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) from a second subset of individuals in the same reference population that have received treatment with the non-PD-L1 axis binding antagonist therapy that does not contain the PD-L1 axis binding antagonist, wherein the responsiveness of the individual to treatment with the non-PD-L1 axis binding antagonist therapy is significantly (e.g., statistically significantly) improved relative to the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist.

The reference immune score expression level can be determined from any number of individuals in a reference population and/or any number of reference samples (e.g., reference cells, reference tissues, control samples, control cells, or control tissues). The reference sample may be a single sample or a combination of multiple samples. Reference immune score expression levels based on reference samples can be based on any number of reference samples (e.g., 2 or more, 5 or more, 10 or more, 50 or more, 100 or more, 500 or more, or 1000 or more reference samples). In some cases, the reference sample comprises pooled mRNA samples extracted from samples obtained from multiple individuals. Further, a reference immune score expression level based on a reference population or sample thereof can be based on any number of individuals in the reference population (e.g., 2 or more, 5 or more, 10 or more, 50 or more, 100 or more, 500 or more, or 1000 or more individuals in the reference population). The reference immune score expression level can be determined from the measurements based on multiple samples or multiple individuals in the reference population using any statistical method known in the art. See, for example: sokal R.R. and Rholf, F.J. (1995) "Biometry: the principles and practice of statistics in biological research", W.H.Freeman and Co., N.Y..

(viii) Reference population

The reference immune score expression level may reflect one or more expression levels of one or more genes described herein (e.g., one or more genes listed in any one of tables 1-17) in one or more reference populations (or reference samples), or a pre-specified reference value.

In some cases, the reference immune score expression level is an immune score expression level of one or more genes listed in any one of tables 1 to 17 in the reference population.

In some cases, the reference population is a population of individuals with cancer. In some cases, the reference population is a population of individuals with lung cancer (e.g., NSCLC). In some cases, the reference population is a population of individuals suffering from kidney cancer (e.g., RCC). In some cases, the reference population is a population of individuals with bladder cancer (e.g., UC). In some cases, the reference population is a population of individuals with breast cancer (e.g., TNBC). In some cases, the reference population is a population of individuals not suffering from cancer.

Further, the reference population can include one or more subsets (e.g., one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or ten or more subsets) of individuals.

In some cases, the reference population is a population of individuals having cancer, wherein the population of individuals comprises a subset of individuals that have received treatment with at least one dose (e.g., at least one dose, at least two doses, at least three doses, at least four doses, at least five doses, at least six doses, at least seven doses, at least eight doses, at least nine doses, or at least ten doses, or more than ten doses) of therapy including a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)). In some cases, a therapy comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) is a monotherapy. In other instances, the therapy comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) is a combination therapy comprising at least one additional therapeutic agent (e.g., an anti-cancer therapy (e.g., an anti-tumor agent, a chemotherapeutic agent, a growth inhibitory agent, a cytotoxic agent, radiation therapy, or a combination thereof)) in addition to the PD-L1 axis binding antagonist.

In some cases, the reference population is a population of individuals with cancer, wherein the population of individuals includes a population of individuals that have received treatment with therapy that does not include a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) non-PD-L1 axis binding antagonist therapy (e.g., an anti-cancer therapy (e.g., an anti-tumor agent, a chemotherapeutic agent, a growth inhibitory agent, a cytotoxic agent, radiation therapy, or a combination thereof)).

In some cases, the reference population includes a combination of individuals from different subsets. For example, in some cases, the reference population may be a population of individuals with cancer, including: (i) A first subset of individuals who have received treatment with a PD-L1 axis binding antagonist therapy (e.g., PD-L1 binding antagonist therapy) comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)); and (ii) a second subset of individuals treated with a non-PD-L1 axis binding antagonist therapy (e.g., a non-PD-L1 binding antagonist therapy) that does not include a PD-L1 axis binding antagonist (e.g., an anti-cancer therapy (e.g., an anti-tumor agent, a chemotherapeutic agent, a growth inhibitory agent, a cytotoxic agent, radiation therapy, or a combination thereof).

Methods of treatment and therapeutic uses

Also provided herein are methods, medicaments, and uses thereof for treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., alemtuzumab), or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) based on the presence and/or expression level of one or more of any one of the biomarkers disclosed herein having been measured in a sample from the individual.

Any of the methods provided herein can comprise determining the presence and/or expression level of any of the biomarkers disclosed herein. In some cases, any of the methods provided herein can comprise administering a PD-L1 binding antagonist to an individual based on a previously determined presence and/or expression level of any of the biomarkers disclosed herein. In other instances, any of the methods provided herein can comprise administering a PD-L1 binding antagonist to an individual prior to determining the presence and/or expression level of any of the biomarkers disclosed herein.

Any of the methods, medicaments, and uses can include or involve any of the PD-L1 axis binding antagonists disclosed herein (e.g., in section IV).

For example, a biomarker may comprise the presence and/or level of expression of a biomarker listed in any one of tables 1 to 17 in a sample obtained from an individual; the presence of TLS in a sample obtained from an individual; the number of B cells in a sample obtained from the individual; the presence of clonally expanded B cells in a sample from an individual; and/or combinations thereof. Any suitable sample may be used, for example, any sample type disclosed herein, including tumor samples.

For example, provided herein is a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), renal cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) genes listed in table 1 in a sample from the individual, wherein the immune score expression level of the one or more genes in the sample is determined to be higher than a reference immune score expression level of the two or more genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, provided herein is a method of treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have an immune score expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) genes listed in table 1 in a sample from the individual that is higher than a reference immune score expression level of the one or more genes, comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., an anti-PD-L1 antibody, such as attrituximab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

Any suitable immuno-score reference expression level can be used. In some cases, the immune score reference expression level is an immune score expression level of one or more genes in a reference population. In some cases, the reference population is a population of individuals with cancer. In some cases, the population of individuals includes a first subset of individuals who have received treatment with a PD-L1 axis binding antagonist and a second subset of individuals who have received treatment with a therapy that does not contain a PD-L1 axis binding antagonist. In some cases, the reference immune score expression level visibly distinguishes between the first subset of individuals and each of the second subset of individuals based on a significant difference between the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist and the responsiveness of the individual to treatment with the therapy that does not contain the PD-L1 axis binding antagonist above the reference immune score expression level, wherein the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist is significantly improved relative to the responsiveness of the individual to treatment with the therapy that does not contain the PD-L1 axis binding antagonist. In some cases, the therapy without the PD-L1 axis binding antagonist comprises an anti-tumor agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, or a combination thereof. In some cases, the therapy without the PD-L1 axis binding antagonist comprises a chemotherapeutic agent. In some cases, the chemotherapeutic agent is docetaxel. In some cases, responsiveness to treatment comprises prolongation of OS, prolongation of progression-free survival (PFS), or an increase in the determined optimal overall response (BCOR). In some cases, responsiveness to treatment includes an extension of Overall Survival (OS). In some cases, the reference immune score expression level is the median of the expression levels of each of the one or more genes in the reference population. In some cases, the median expression level is the median of the mean Z scores for the expression levels of each of the one or more genes in the reference population.

The cancer may be any suitable type of cancer. In some cases, the cancer is lung cancer, kidney cancer, bladder cancer, breast cancer, colorectal cancer, ovarian cancer, pancreatic cancer, gastric cancer, esophageal cancer, mesothelioma, melanoma, head and neck cancer, thyroid cancer, sarcoma, prostate cancer, glioblastoma, cervical cancer, thymus cancer, leukemia, lymphoma, myeloma, mycosis fungoides, merkel cell carcinoma, or hematological malignancy. In some cases, the cancer is lung, kidney, bladder, or breast cancer. In some cases, the lung cancer is non-small cell lung cancer (NSCLC). In some cases, the NSCLC is non-squamous NSCLC. In some cases, the NSCLC is squamous NSCLC.

A.B cell characteristics

(i) Gene signature associated with B cells

In some aspects, the methods provided herein can involve administering a PD-L1 axis binding antagonist to an individual based on the expression level of one or more genes in the B cell signature. Any suitable B cell characteristics may be used. For example, a B cell characteristic can include one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) genes listed in table 2.

For example, provided herein is a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), renal cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein the immune score expression level of the one or more genes in the sample is determined to be higher than a reference immune score expression level of the one or more genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, provided herein is a method of treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have an immune score expression level of one or more of genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) in a sample from the individual that is higher than a reference immune score expression level of the one or more genes, the method comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attritol) or a PD-1 binding antagonist (e.g., anti-PD 1 antibody)).

In yet another example, provided herein is a PD-L1 axis binding antagonist for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein the immune score expression level of the one or more genes in the sample is determined to be higher than a reference immune score expression level of the one or more genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, provided herein is a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for use in treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have an immune score expression level of one or more of the genes CD79A, CD19, BANK1, jcain, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) in a sample from the individual that is higher than a reference immune score expression level of the one or more genes.

In yet another example, provided herein is use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) comprising: (a) Determining the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein the immune score expression level of the one or more genes in the sample is determined to be higher than a reference immune score expression level of the one or more genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another example, also provided herein is the use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have one or more of the genes CD79A, CD19, BANK1, jcain, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) in a sample from the individual at an immune expression level that is greater than a reference immune score for the one or more genes.

In some cases, the method comprises determining the expression level of one of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB 1.

In some cases, the method comprises determining the expression level of CD 79A.

In some cases, the method comprises determining the expression level of CD 19.

In some cases, the method comprises determining the expression level of BANK 1.

In some cases, the method comprises determining the expression level of JCHAIN.

In some cases, the method includes determining the expression level of SLAMF 7.

In some cases, the method includes determining an expression level of BTK.

In some cases, the method comprises determining the expression level of IGJ.

In some cases, the method includes determining the expression level of IGLL 5.

In some cases, the method comprises determining the expression level of RBPJ.

In some cases, the method comprises determining an expression level of MZB 1.

In some cases, the expression level of one of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 has been determined.

In some cases, the expression level of CD79A has been determined.

In some cases, the expression level of CD19 has been determined.

In some cases, the expression level of BANK1 has been determined.

In some cases, the expression level of JCHAIN has been determined.

In some cases, the expression level of SLAMF7 has been determined.

In some cases, the expression level of BTK has been determined.

In some cases, the expression level of TNFRSF17 has been determined.

In some cases, the expression level of IGJ has been determined.

In some cases, the expression level of IGLL5 has been determined.

In some cases, the expression level of RBPJ has been determined.

In some cases, the expression level of MZB1 has been determined.

For example, provided herein is a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining in a sample from the individual the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1, wherein determining the immune score expression level of the one or more genes in the sample is greater than a reference immune score expression level of the one or more genes identifies the individual as an individual likely to benefit from treatment comprising a PD-L1 axis binding antagonist, wherein the benefit comprises a prolonged OS in the individual as compared to treatment without the PD-L1 axis binding antagonist; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, provided herein is a method of treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have an immune score expression level of one or more of genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, rbj, and MZB1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) in a sample from the individual that is higher than a reference immune score expression level of the one or more genes, the method comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attritrin antagonist) or a PD-1 binding antagonist (e.g., a PD-1 axis binding antagonist), wherein the increased immune score expression level of the individual is greater than the reference immune score of the reference gene, wherein the individual's increased immune system comprising the use of the one or more of the individual.

In yet another example, provided herein is a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining in a sample from the individual the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1, wherein determining the immune score expression level of the one or more genes in the sample is greater than a reference immune score expression level of the one or more genes identifies the individual as an individual likely to benefit from treatment comprising a PD-L1 axis binding antagonist, wherein the benefit comprises a prolonged OS in the individual as compared to treatment without the PD-L1 axis binding antagonist; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another example, provided herein is a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for use in treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have an immune score for one or more of genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) in a sample from the individual that is higher than a reference immune score for the individual that would benefit from the use of the reference immune score for the one or more genes, wherein the individual has an immune score that is higher than a reference level of expression of the individual that would benefit from the increased expression of the one or more genes.

In yet another example, provided herein is use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) comprising: (a) Determining in a sample from the individual the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1, wherein determining the immune score expression level of the one or more genes in the sample is greater than a reference immune score expression level of the one or more genes identifies the individual as an individual likely to benefit from treatment comprising a PD-L1 axis binding antagonist, wherein the benefit comprises a prolonged OS in the individual as compared to treatment without the PD-L1 axis binding antagonist; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another example, also provided herein is the use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have one or more of the genes CD79A, CD19, BANK1, jcha in, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) in a sample from the individual, wherein the individual has an immune expression score that is higher than a reference immune expression score for the one or more genes, wherein the identification of the subject's immune expression score is likely to benefit from use of the PD-L1 binding antagonist.

For example, provided herein is a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining in a sample from the individual the expression levels of two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1, wherein the two or more genes in the sample are determined to have an immune score expression level that is higher than a reference immune score expression level for the two or more genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, provided herein is a method of treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have an immune score expression level of two or more of genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, rbj, and MZB1 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) in a sample from the individual that is higher than a reference immune score expression level of the two or more genes, the method comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attritol monoclonal antibody) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In yet another example, provided herein is a PD-L1 axis binding antagonist for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining in a sample from the individual the expression level of two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1, wherein the determination of the immune score expression level of the two or more genes in the sample is higher than the reference immune score expression level of the two or more genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, provided herein is a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for use in treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have an immune expression level of two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB 1) in a sample from the individual that is higher than a reference immune score for the two or more genes.

In yet another example, provided herein is use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) comprising: (a) Determining the expression level of two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein the immune score expression level of the two or more genes in the sample is determined to be higher than a reference immune score expression level of the two or more genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another example, also provided herein is the use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have an immune score expression level of two or more of the genes CD79A, CD19, BANK1, jcain, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual that is higher than a reference immune expression level of the two or more genes.

Any combination of B cell signature genes can be determined. For example, the combination may include two genes selected from CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1, e.g., any one of the combinations listed in table 3. In another example, the combination can include three genes selected from CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1, e.g., any one of the combinations listed in table 4. In another example, the combination can include four genes selected from CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1, e.g., any one of the combinations listed in table 5. In another example, the combination can include five genes selected from CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1, e.g., any one of the combinations listed in table 6. In another example, the combination can include six genes selected from CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1, e.g., any one of the combinations listed in table 7. In another example, the combination may include seven genes selected from CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1, e.g., any one of the combinations listed in table 8.

(ii) Gene signature associated with plasma B cells

In some aspects, the methods provided herein can involve administering a PD-L1 axis binding antagonist to an individual based on the expression level of one or more genes in a plasma B cell signature. Any suitable plasma B cell characteristics may be used. For example, a plasma B cell characteristic can include one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14) genes listed in table 9.

For example, provided herein is a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining in a sample from the individual the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14) of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5, wherein the determination of the immune expression level of the one or more genes in the sample is higher than the reference immune score expression level of the one or more genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, provided herein is a method of treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have an immune expression level of one or more of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14) in a sample from the individual that is higher than a reference immune expression level of the one or more genes, the method comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., binding to an anti-PD antibody (e.g., attorneol 1-PD), such as an anti-PD 1-PD antibody).

In yet another example, provided herein is a PD-L1 axis binding antagonist for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining in a sample from the individual the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14) of the genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5, wherein the determination of the immune expression level of the one or more genes in the sample is higher than the reference immune score expression level of the one or more genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, provided herein is a PD-L1 axis binding antagonist (e.g., PD-L1 binding antagonist (e.g., anti-PD-L1 antibody, e.g., altlizumab)) for use in treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have one or more of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14) in a sample from the individual at a high immune score or at one or more immune expression level of the immune score.

In yet another example, provided herein is the use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., NSCLC), renal cancer (e.g., rcu), and breast cancer (e.g., TNBC)) comprising: (a) Determining in a sample from the individual the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14) of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5, wherein the determination of the immune expression level of the one or more genes in the sample is higher than the reference immune score expression level of the one or more genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another example, also provided herein is the use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attrituzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have one or more of the genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 (e.g., one or more of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, or more immune scores) in a sample from the individual at a high immune expression level of the reference.

In some cases, the method comprises determining an expression level of MZB 1.

In some cases, the method comprises determining an expression level of DERL 3.

In some cases, the method comprises determining an expression level of JSRP 1.

In some cases, the method comprises determining the expression level of IGHG 2.

In some cases, the method comprises determining the expression level of IGHGP.

In some cases, the method comprises determining the expression level of IGHA 2.

In some cases, the method comprises determining the expression level of IGLC 7.

In some cases, the method includes determining the expression level of IGLL 5.

In some cases, the expression level of one of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 has been determined.

In some cases, the expression level of MZB1 has been determined.

In some cases, the expression level of DERL3 has been determined.

In some cases, the expression level of JSRP1 has been determined.

In some cases, the expression level of TNFRSF17 has been determined.

In some cases, the expression level of SLAMF7 has been determined.

In some cases, the expression level of IGHG2 has been determined.

In some cases, the expression level of IGHGP has been determined.

In some cases, the expression level of IGLV3-1 has been determined.

In some cases, the expression level of IGLV6-57 has been determined.

In some cases, the expression level of IGHA2 has been determined.

In some cases, the expression level of IGKV4-1 has been determined.

In some cases, the expression level of IGKV1-12 has been determined.

In some cases, the expression level of IGLC7 has been determined.

In some cases, the expression level of IGLL5 has been determined.

For example, provided herein is a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining in a sample from the individual the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14) of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5, wherein determining the immune expression level of the one or more genes in the sample is greater than a reference immune score expression level for the one or more genes identifies the individual as likely to comprise treatment with a PD-L1 axis binding antagonist, wherein the benefit comprises a prolongation of the individual's OS as compared to treatment without a PD-L1 axis binding antagonist; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, provided herein is a method of treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have an immune expression score for one or more of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14) in a sample from the individual that is higher than a reference immune score expression level for the one or more genes, the method comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) wherein in the event the immune score expression level of the one or more genes is greater than the reference immune score expression level of the one or more genes, identifying the individual as an individual who is likely to benefit from treatment comprising the PD-L1 axis binding antagonist, wherein the benefit comprises a prolongation of OS in the individual as compared to treatment without the PD-L1 axis binding antagonist.

In yet another example, provided herein is a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining in a sample from the individual the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14) of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5, wherein determining the immune expression level of the one or more genes in the sample is greater than a reference immune score expression level for the one or more genes identifies the individual as an individual likely to comprise treatment with a PD-L1 axis binding antagonist, wherein the benefit comprises a prolongation of OS in the individual as compared to treatment without the PD-L1 axis binding antagonist; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another example, provided herein is a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have one or more of the genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 13) in a sample from the individual as having an immune response to a high level of immune expression of the gene expression, wherein the individual comprises a reference level of the immune-L expression of the immune antagonist that would benefit from treatment or a reference level of the individual.

In yet another example, provided herein is use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) comprising: (a) Determining in a sample from the individual the expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14) of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5, wherein determining the immune expression level of the one or more genes in the sample is greater than a reference immune score expression level for the one or more genes identifies the individual as an individual likely to comprise treatment with a PD-L1 axis binding antagonist, wherein the benefit comprises a prolongation of OS in the individual as compared to treatment without the PD-L1 axis binding antagonist; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another example, also provided herein is the use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as alemtuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for treating a cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have one or more of the genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV 4-1-12, IGLC7, and IGLL5 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or IGLV 1-12) in a sample from the individual, wherein the individual has an expression of the gene at a level that is higher than a reference level that would benefit from the use of the one or more of the PD-L1-1 expression of the PD-1, or 11 gene expression of the individual.

B.TLS

In some aspects, the methods provided herein can involve administering a PD-L1 axis binding antagonist to an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) based on the presence of TLS in a sample (e.g., tumor sample) from the individual.

For example, provided herein is a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining the presence of TLS in a sample (e.g., a tumor sample) from the individual; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, provided herein is a method of treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have TLS present in a sample (e.g., a tumor sample) from the individual, the method comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In yet another example, provided herein is a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining the presence of TLS in a sample (e.g., a tumor sample) from the individual; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another example, provided herein is a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for use in treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have TLS present in a sample (e.g., a tumor sample) from the individual.

In yet another example, provided herein is the use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., NSCLC), renal cancer (e.g., rcu), and breast cancer (e.g., TNBC)) comprising: (a) Determining the presence of TLS in a sample (e.g., a tumor sample) from the individual; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another example, also provided herein is the use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attritumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have TLS present in a sample (e.g., a tumor sample) from the individual.

Any suitable histological staining method may be used. For example, in some cases, histological staining includes H & E staining.

Any suitable IHC or immunofluorescence method may be used. In some cases, IHC or immunofluorescence includes detecting CD62L, L-selectin, CD40, or CD8, e.g., using antibodies (e.g., anti-CD 62L antibodies, anti-L-selectin antibodies, anti-CD 40 antibodies, and/or anti-CD 8 antibodies). In some cases, CD62L or L-selectin is detected using a MECA-79 antibody.

In some cases, the sample may include one TLS. In other cases, a sample can include more than one TLS, e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 50, 60, 70, 80, 90, 100, or more TLS.

C.TLS feature

In some aspects, the methods provided herein can involve administering a PD-L1 axis binding antagonist to an individual based on the expression level of one or more genes in the TLS signature. Any suitable TLS feature may be used. For example, the TLS signature can include one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) genes listed in table 16.

For example, provided herein is a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining an expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual, wherein the determination of the immune score expression level of the one or more genes in the sample is higher than a reference immune score expression level of the one or more genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, provided herein is a method of treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have an immune score expression level of one or more of genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) in a sample from the individual that is higher than a reference immune score expression level of the one or more genes, the method comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attritrin bead) or a PD-1 binding antagonist (e.g., anti-PD 1 antibody)).

In yet another example, provided herein is a PD-L1 axis binding antagonist for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining an expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual, wherein the determination of the immune score expression level of the one or more genes in the sample is higher than a reference immune score expression level of the one or more genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, provided herein is a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for use in treating a cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have one or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) in a sample from the individual at an immune score level that is higher than a reference immune score level for the one or more of the one or more genes.

In another example, provided herein is the use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) comprising: (a) Determining an expression level of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual, wherein determining an immune score expression level of the one or more genes in the sample is higher than a reference immune score expression level of the one or more genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another example, also provided herein is the use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attritumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for treating a cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have one or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) in a sample from the individual at an immune expression level that is greater than a reference immune expression level for the one or more genes.

For example, provided herein is a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining an expression level of two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual, wherein determining an immune score expression level of the two or more genes in the sample is higher than a reference immune score expression level of the two or more genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, provided herein is a method of treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have an immune score expression level of two or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) in a sample from the individual that is higher than a reference immune score expression level of the two or more genes, the method comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attritrin monoclonal antibody) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In yet another example, provided herein is a PD-L1 axis binding antagonist for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), renal cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining an expression level of two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual, wherein determining an immune score expression level of the two or more genes in the sample is higher than a reference immune score expression level of the two or more genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, provided herein is a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attrituximab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for use in treating a cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have an immune score expression level of two or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) in a sample from the individual that is higher than a reference immune score expression level of the two or more genes.

In another example, provided herein is the use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) comprising: (a) Determining an expression level of two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual, wherein determining an immune score expression level of the two or more genes in the sample is higher than a reference immune score expression level of the two or more genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another example, also provided herein is the use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attritumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for treating a cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have an immune score expression level of two or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) in a sample from the individual that is higher than a reference immune expression level of the two or more genes.

Any combination of TLS signature genes can be identified, for example, as any combination of two genes selected from: CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13; any combination of three genes selected from: CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13; any combination of four genes selected from: CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13; any combination of five genes selected from: CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13; any combination of six genes selected from: CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13; any combination of seven genes selected from: CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13; any combination of eight genes selected from: CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13; any combination of nine genes selected from: CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13; any combination of ten genes selected from: CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13; any combination of eleven genes selected from: CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13; or any combination of eleven genes selected from: CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11 and CXCL13.

B cell number and clonally expanded B cells

In some aspects, the methods provided herein can involve administering a PD-L1 axis binding antagonist to an individual based on the presence and/or number of B cells in a sample from the individual. In some aspects, the methods provided herein can involve determining the presence and/or number of clonally expanded B cells in a sample from an individual.

For example, provided herein is a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining the number of B cells in a tumor sample from the individual; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, provided herein is a method of treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have a number of B cells in a tumor sample from the individual that is greater than a reference number of B cells, the method comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In yet another example, provided herein is a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining the number of B cells in a tumor sample from the individual; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another example, provided herein is a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody) for treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have a number of B cells in a tumor sample from the individual that is higher than a reference number of B cells.

In another example, provided herein is the use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) comprising: (a) Determining the number of B cells in a tumor sample from the individual; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another example, also provided herein is the use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentimab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have a number of B cells in a tumor sample from the individual that is higher than a reference number of B cells.

The presence and/or number of any suitable type of B cells can be determined. For example, in some cases, B cells include CD79+ B cells, igG + B cells, and/or plasma cells.

For example, provided herein is a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), renal cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining that the individual has clonally expanded B cells in a tumor sample from the individual; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, provided herein is a method of treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have clonally expanded B cells in a tumor sample from the individual, the method comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In yet another example, provided herein is a PD-L1 axis binding antagonist for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), renal cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining that the individual has clonally expanded B cells in a tumor sample from the individual; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, provided herein is a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody) for treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have clonally expanded B cells in a tumor sample from the individual.

In another example, provided herein is the use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) comprising: (a) Determining that the individual has clonally expanded B cells in a tumor sample from the individual; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist.

In another example, also provided herein is the use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have clonally expanded B cells in a tumor sample from the individual.

The clonally expanded B cells may be any type of B cell. For example, in some cases, the clonally expanded B cells are clonally expanded plasma cells.

E.T Effector characteristics

In some aspects, the methods provided herein can involve administering a PD-L1 axis binding antagonist to an individual based on the presence and/or expression level of one or more T effector feature genes. Any suitable T effector feature may be used. For example, the T effector characteristic may include one or more (e.g., 1, 2, 3, 4, 5, 6, 7, or 8) genes listed in table 17. In some cases, CD274 is further detected.

For example, provided herein is a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining in a sample from the individual the expression levels of (i) one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) of the genes listed in table 1 and (ii) one or more (e.g., 1, 2, 3, 4, 5, 6, 7, or 8) of the genes CD8A, EOMES, GZMA, TBX21, IFNG, GZMB, CXCL9, and CXCL10, wherein the one or more genes listed in (i) table 1 and (ii) the genes CD8A, EOMES, GZMB a, TBX21, IFNG, GZMB, CXCL9, and CXCL10 in the sample are determined to have a high expression score in the immune score of the one or more genes listed in (i) table 1, the reference gene or genes CD 21, the reference gene score of (i, cxmes, CXCL9, ifg 2, 5, 6, 7, or 8); and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In yet another example, provided herein is a PD-L1 axis binding antagonist for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), renal cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining in a sample from the individual the expression levels of (i) one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) of the genes listed in table 1 and (ii) one or more (e.g., 1, 2, 3, 4, 5, 6, 7, or 8) of the genes CD8A, EOMES, GZMA, TBX21, IFNG, GZMB, CXCL9, and CXCL10, wherein the one or more genes listed in (i) table 1 and (ii) the genes CD8A, EOMES, GZMB a, TBX21, IFNG, GZMB, CXCL9, and CXCL10 in the sample are determined to have a high expression score in the immune score of the one or more genes listed in (i) table 1, the reference gene or genes CD 21, the reference gene score of (i, cxmes, CXCL9, ifg 2, 5, 6, 7, or 8); and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In yet another example, provided herein is the use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., NSCLC), renal cancer (e.g., rcu), and breast cancer (e.g., TNBC)) comprising: (a) Determining in a sample from the subject the expression levels of (i) one or more of the genes listed in table 1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20) and (ii) one or more of the genes CD8A, EOMES, GZMA, TBX21, IFNG, GZMB, CXCL9 and CXCL10 (e.g., 1, 2, 3, 4, 5, 6, 7 or 8), wherein the expression levels of (i) one or more of the genes listed in table 1 and (ii) the genes CD8A, EOMES, GZMB a, TBX21, IFNG, GZMB, CXCL9 and CXCL10 in the sample are determined to be higher than the expression levels of (i) one or more of the genes listed in table 1 and (ii) the immune score of the genes CD8A, cxme, CXCL 21, cxsa, CXCL9 and CD 10; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, provided herein is a method of treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have in a sample from the individual one or more of (i) one or more of the genes listed in table 1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) and (ii) one or more of the genes CD8A, EOMES, GZMA, TBX21, IFNG, gzm b, CXCL9, and CXCL10 (e.g., 1, 2, 3, 4, 5, 6, 7, or 8) (e.g., the level of immune expression of the antagonist is higher than the level of the anti-CXCL 1, e.g., anti-CD 1, 2, 3, 4, 5, 6, 7, or 8) antagonists listed in the individual (e.g., binding to the anti-CXCL 1, e.g., anti-CD 1, 2, 3, 4, or 1, e.g., binding to the anti-spg., anti-CD 1, or anti-CD 1) gene).

In another example, provided herein is a PD-L1 axis binding antagonist (e.g., PD-L1 binding antagonist (e.g., anti-PD-L1 antibody, e.g., attrituzumab) or PD-1 binding antagonist (e.g., anti-PD-1 antibody)) for use in treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have in a sample from the individual a high level of expression of one or more of (i) the genes listed in table 1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) and (ii) one or more of (e.g., 5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) of the genes listed in (ii) CD8A, EOMES, gcmxa, gcmx, gccxa, gcx 21, ifgcl, and (ifgcl) genes.

In another example, also provided herein is the use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have in a sample from the individual (i) one or more of the genes listed in table 1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) and (ii) one or more (e.g., 1, 2, 3, 4, 5, 6, 7, or 8) of the genes CD8A, EOMES, GZMA, TBX21, IFNG, GZMB, CXCL9, and CXCL10, is higher than the reference immune score expression level of (i) one or more of the genes listed in table 1 and (ii) one or more of the genes CD8A, EOMES, GZMA, TBX21, IFNG, GZMB, CXCL9, and CXCL 10.

In some cases, the reference immune score expression level is an immune score expression level of one or more of (i) one or more genes listed in table 1 and (ii) one or more of the genes CD8A, EOMES, GZMA, TBX21, IFNG, GZMB, CXCL9, and CXCL10 in the reference population. In some cases, the reference population is a population of individuals with cancer. In some cases, the population of individuals includes a first subset of individuals who have received treatment with a PD-L1 axis binding antagonist and a second subset of individuals who have received treatment with a therapy that does not contain a PD-L1 axis binding antagonist. In some cases, the reference immune score expression level clearly distinguishes each of the first subset of individuals from the second subset of individuals based on a significant difference between the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist and the responsiveness of the individual to treatment with the therapy that does not contain the PD-L1 axis binding antagonist above the reference expression level, where the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist is significantly improved relative to the responsiveness of the individual to treatment with the therapy that does not contain the PD-L1 axis binding antagonist. In some cases, the therapy without the PD-L1 axis binding antagonist comprises an anti-neoplastic agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, or a combination thereof. In some cases, the therapy without the PD-L1 axis binding antagonist comprises a chemotherapeutic agent. In some cases, the chemotherapeutic agent is docetaxel. In some cases, responsiveness to treatment comprises prolongation of OS, prolongation of PFS, or an increase in BCOR. In some cases, responsiveness to treatment includes an extension of Overall Survival (OS). In some cases, the reference immune score expression level is the median of the expression levels of each of the two or more genes in the reference population. In some cases, the median expression level is the median of the average Z-scores for the expression levels of each of (i) the one or more genes listed in table 1 and (ii) the genes CD8A, EOMES, GZMA, TBX21, IFNG, GZMB, CXCL9, and CXCL10 in the reference population.

For example, provided herein is a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining in a sample from the individual the expression levels of (i) one or more of the B cell characteristic genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ and MZB1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11) and (ii) one or more of the T effector characteristic genes CD8A, EOMES, GZMA, TBX21, IFNG, GZMB, CXCL9 and CXCL10 (e.g., 1, 2, 3, 4, 5, 6, 7 or 8), wherein the determination of the immune expression levels of (i) the one or more B cell characteristic genes and (ii) the one or more T effector characteristic genes in the sample is greater than the immune score of (i) the one or more B cell characteristic genes and (ii) the one or more T effector characteristic genes and the reference expression level of (i) the one or more T effector characteristic genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, provided herein is a method of treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have in a sample from the individual one or more of (i) the B cell signature genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, rbzpj, and MZB1 (e.g., one or more of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) and (ii) the T effector signature genes CD8A, mes, ga, TBX21, IFNG, GZMB, CXCL9, and CXCL10 (e.g., 1, 2, 3, 4, 5, 6, 7, or 8) is higher than a reference immune score expression level of (i) the one or more B cell signature genes and (ii) the one or more T effector signature genes, the method comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as astuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In yet another example, provided herein is a PD-L1 axis binding antagonist for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining in a sample from the individual the expression levels of (i) one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) of B cell characteristic genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 and (ii) one or more (e.g., 1, 2, 3, 4, 5, 6, 7, or 8) T effector characteristic genes CD8A, EOMES, GZMA, TBX21, ifb, GZMB, CXCL9, and CXCL10, wherein the determination of the expression levels of (i) the one or more B cell characteristic genes and (ii) the one or more T effector characteristic genes in the sample is greater than the immune score of (i) the one or more B cell characteristic genes and (ii) the one or more T effector characteristic genes is greater than the immune score of (i) of the one or more B cell characteristic genes and (ii) the immune effector characteristic genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, provided herein is a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for use in treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have in a sample from the individual one or more of (i) B cell signature genes CD79A, CD19, BANK1, jcain, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) and (ii) one or more (e.g., 1, 2, 3, 4, 5, 6, 7, or 8) of the T effector signature genes CD8A, EOMES, GZMA, TBX21, IFNG, GZMB, CXCL9, and CXCL10 are higher than the reference immune score expression level of (i) the one or more B cell signature genes and (ii) the one or more T effector signature genes.

In yet another example, provided herein is the use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., NSCLC), renal cancer (e.g., rcu), and breast cancer (e.g., TNBC)) comprising: (a) Determining in a sample from the individual the expression levels of (i) one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) of B cell characteristic genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 and (ii) one or more (e.g., 1, 2, 3, 4, 5, 6, 7, or 8) T effector characteristic genes CD8A, EOMES, GZMA, TBX21, ifb, GZMB, CXCL9, and CXCL10, wherein the determination of the expression levels of (i) the one or more B cell characteristic genes and (ii) the one or more T effector characteristic genes in the sample is greater than the immune score of (i) the one or more B cell characteristic genes and (ii) the one or more T effector characteristic genes is greater than the immune score of (i) of the one or more B cell characteristic genes and (ii) the immune effector characteristic genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, also provided herein is the use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for treating a cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have in a sample from the individual one or more of (i) B cell signature genes CD79A, CD19, BANK1, jcha, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) and (ii) one or more (e.g., 1, 2, 3, 4, 5, 6, 7, or 8) of the T effector characteristic genes CD8A, EOMES, GZMA, TBX21, IFNG, GZMB, CXCL9, and CXCL10, is higher than the reference immune score expression level of (i) the one or more B cell characteristic genes and (ii) the one or more T effector characteristic genes.

In some cases, the reference immune score expression level is an immune score expression level of (i) the one or more B cell signature genes and (ii) the one or more T effector signature genes in the reference population. In some cases, the reference population is a population of individuals with cancer. In some cases, the population of individuals includes a first subset of individuals who have received treatment with a PD-L1 axis binding antagonist and a second subset of individuals who have received treatment with a therapy that does not contain a PD-L1 axis binding antagonist. In some cases, the reference immune score expression level clearly distinguishes each of the first subset of individuals from the second subset of individuals based on a significant difference between the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist and the responsiveness of the individual to treatment with the therapy that does not contain the PD-L1 axis binding antagonist above the reference expression level, where the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist is significantly improved relative to the responsiveness of the individual to treatment with the therapy that does not contain the PD-L1 axis binding antagonist. In some cases, the therapy without the PD-L1 axis binding antagonist comprises an anti-tumor agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, or a combination thereof. In some cases, the therapy without the PD-L1 axis binding antagonist comprises a chemotherapeutic agent. In some cases, the chemotherapeutic agent is docetaxel. In some cases, responsiveness to treatment comprises a prolongation of OS, a prolongation of PFS, or an increase in BCOR. In some cases, responsiveness to treatment includes an extension of Overall Survival (OS). In some cases, the reference immune score expression level is the median of the expression levels of each of the two or more genes in the reference population. In some cases, the median expression level is the median of the mean Z scores for the expression levels of each of (i) the one or more B cell signature genes and (ii) the one or more T effector signature genes in the reference population.

In another example, provided herein is a method of treating cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have in a sample from the individual one or more of (i) the TLS signature genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 (e.g., one or more of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) and (ii) the T effector signature genes CD8A, mes, ga, TBX21, IFNG, GZMB, CXCL9, and CXCL10 (e.g., 1, 2, 3, 4, 5, 6, 7, or 8) is higher than a reference immune score expression level of (i) the one or more TLS signature genes and (ii) the one or more T effector signature genes, the method comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attritumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In yet another example, provided herein is a PD-L1 axis binding antagonist for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)), the method comprising: (a) Determining in a sample from the individual the expression levels of (i) one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) of the TLS signature genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 and (ii) one or more (e.g., 1, 2, 3, 4, 5, 6, 7, or 8) of the T effector signature genes CD8A, EOMES, GZMA, TBX21, ifb, GZMB, CXCL9, and CXCL10, wherein the immune expression levels of (i) the one or more TLS signature genes and (ii) the one or more T effector signature genes in the sample are determined to be higher than the immune expression level of (i) the one or more TLS signature genes and (ii) of the one or more T effector signature genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, provided herein is a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for use in treating a cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have in a sample from the individual one or more of (i) the TLS signature genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) and (ii) one or more (e.g., 1, 2, 3, 4, 5, 6, 7, or 8) of the T effector signature genes CD8A, EOMES, GZMA, TBX21, IFNG, GZMB, CXCL9, and CXCL10 are higher than the reference immune score expression level of (i) the one or more TLS signature genes and (ii) the one or more T effector signature genes.

In yet another example, provided herein is use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for use in a method of treating an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) comprising: (a) Determining in a sample from the individual the expression level of (i) one or more of the TLS signature genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11 and CXCL13 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12) and (ii) one or more of the T effector signature genes CD8A, EOMES, GZMA, TBX21, IFNG, GZMB, CXCL9 and CXCL10 (e.g., 1, 2, 3, 4, 5, 6, 7 or 8), wherein the immune expression level of (i) the one or more TLS signature genes and (ii) the one or more T effector signature genes in the sample is determined to be higher than the immune expression level of (i) the one or more TLS signature genes and (ii) the one or more T effector signature genes and the reference expression score of (i) or more TLS signature genes; and (b) administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, also provided herein is the use of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as astuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) for the manufacture of a medicament for treating a cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) in an individual, wherein the individual has been determined to have in a sample from the individual (i) one or more of TLS signature genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) and (ii) T response signature genes expressed at a, eoa, 5, 7, 8, 9, 10, 11, or 12, or 5, 7, or 8, e.g., more of the immune signature genes expressed at a high level (e.g., at one or more of the reference genes.

In some cases, the reference immune score expression level is the immune score expression level of (i) the one or more TLS signature genes and (ii) the one or more T effector signature genes in the reference population. In some cases, the reference population is a population of individuals with cancer. In some cases, the population of individuals includes a first subset of individuals who have received treatment with a PD-L1 axis binding antagonist and a second subset of individuals who have received treatment with a therapy that does not contain a PD-L1 axis binding antagonist. In some cases, the reference immune score expression level visibly differentiates each of the first subset of individuals from the second subset of individuals based on a significant difference between the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist and the responsiveness of the individual to treatment with the therapy without the PD-L1 axis binding antagonist above the reference expression level, wherein the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist is significantly improved relative to the responsiveness of the individual to treatment with the therapy without the PD-L1 axis binding antagonist. In some cases, the therapy without the PD-L1 axis binding antagonist comprises an anti-tumor agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, or a combination thereof. In some cases, the therapy without the PD-L1 axis binding antagonist comprises a chemotherapeutic agent. In some cases, the chemotherapeutic agent is docetaxel. In some cases, responsiveness to treatment comprises prolongation of OS, prolongation of PFS, or an increase in BCOR. In some cases, responsiveness to treatment includes an extension of Overall Survival (OS). In some cases, the reference immune score expression level is the median of the expression levels of each of the two or more genes in the reference population. In some cases, the median expression level is the median of the mean Z scores for the expression levels of each of (i) the one or more TLS signature genes and (ii) the one or more T effector signature genes in the reference population.

F. Administration of

PD-L1 axis binding antagonists (e.g., PD-L1 binding antagonists (e.g., anti-PD-L1 antibodies, such as attrituximab) or PD-1 binding antagonists (e.g., anti-PD-1 antibodies)) or compositions thereof and/or any additional therapeutic agents for use in the methods, uses, assays, and kits described herein may be formulated for administration or administered by any suitable method, including, for example: intravenous, intramuscular, subcutaneous, intradermal, transdermal, intraarterial, intraperitoneal, intralesional, intracranial, intraarticular, intraprostatic, intrapleural, intratracheal, intrathecal, intranasal, intravaginal, intrarectal, topical, intratumoral, intraperitoneal, subconjunctival, intracapsular, mucosal, intrapericardial, intraumbilical, intraocular, intraorbital, oral, topical, transdermal, intravitreal (e.g., by intravitreal injection), by eye drop, by inhalation, by injection, by infusion, by continuous infusion, by direct bathing of target cells by local infusion, by catheter, by lavage, by cream, or in a lipid composition. The compositions used in the methods described herein may also be administered systemically or locally. The method of administration may vary depending on a variety of factors (e.g., the compound or composition to be administered and the severity of the condition, disease or disease to be treated). In some cases, the PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., altlizumab) is administered intravenously, intramuscularly, subcutaneously, topically, orally, transdermally, intraperitoneally, intraorbitally, by implantation, by inhalation, intrathecally, intraventricularly, or intranasally. Administration may be by any suitable route, for example by injection, such as intravenous or subcutaneous injection, depending in part on whether administration is transient or chronic. Various dosing schedules are contemplated herein, including but not limited to single or multiple administrations at various time points, bolus administrations, and pulsed infusions.

The PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atelizumab) and any additional therapeutic agents can be formulated, dosed, and administered in a manner consistent with good medical practice. Factors to be considered in this context include the particular condition being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the condition, the site of delivery of the agent, the method of administration, the timing of administration, and other factors known to the practitioner. PD-L1 axis binding antagonists (e.g., PD-L1 binding antagonists, e.g., anti-PD-L1 antibodies, e.g., atelizumab) are formulated and/or administered without, but optionally simultaneously with, one or more drugs currently used to prevent or treat the disorder in question. The effective amount of such other drugs depends on the amount of PD-L1 axis binding antagonist (e.g., PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atlizumab) present in the formulation, the type of disorder or treatment, and other factors described above. These are generally used at the same dosages and routes of administration as described herein, or at about 1% to 99% of the dosages described herein, or at any dosage and by any route empirically/clinically determined to be appropriate.

For the prevention or treatment of cancer (e.g., lung cancer (NSCLC), bladder cancer (UC), kidney cancer (RCC), or breast cancer (TNBC)), the appropriate dosage of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., attritumab) described herein (when used alone or in combination with one or more additional therapeutic agents) will depend on the type of disease to be treated, the severity and course of the disease, the PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., attritumab, administered for either prophylactic or therapeutic purposes, previous treatment, the clinical history of the patient, and the amount of the PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attritumab) or a PD-1 binding antagonist (e.g., anti-PD-1 antibody)), and the discretion of the attending physician, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., attritumab) is suitably administered to the patient at one time or in a series of treatments. Such that the patient receives about two to about twenty, or, for example, about six doses of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attritumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody))). An initial higher loading dose may be administered followed by one or more lower doses. However, other dosage regimens may be useful. The progress of this therapy is readily monitored by conventional techniques and assays.

In some cases, an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) may be between about 60mg to about 5000mg (e.g., between about 60mg to about 4500mg, between about 60mg to about 4000mg, between about 60mg to about 3500mg, between about 60mg to about 3000mg, between about 60mg to about 2500mg, between about 650mg to about 2000mg, between about 60mg to about 1500mg, between about 100mg to about 1500mg, between about 300mg to about 1500mg, between about 500mg to about 1500mg, between about 700mg to about 1500mg, between about 1000mg to about 1000mg, between about 1000mg to about 1400mg, between about 1100mg to about 1300mg, between about 1150mg to about 1250mg, between about 5mg to about 1500mg, between about 1250mg to about 1200mg, or between about 1200.1200 mg ± 2mg, about 1200mg, or about 1200.1200 mg ± 0.1200 mg, e.g. ± 1mg ± 0 mg. In some cases, the method comprises administering to the individual about 1200mg (e.g., a fixed dose of about 1200mg or about 15 mg/kg) of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In some cases, the amount of PD-L1 axis binding antagonist (e.g., PD-L1 binding antagonist (e.g., anti-PD-L1 antibody, e.g., attrituximab) or PD-1 binding antagonist (e.g., anti-PD-1 antibody)) administered to an individual (e.g., a human) can be in the range of 0.01mg/kg body weight of the individual to about 50mg/kg body weight of the individual (e.g., between about 0.01mg/kg and about 45mg/kg, between about 0.01mg/kg and about 40mg/kg, between about 0.01mg/kg and about 35mg/kg, between about 0.01mg/kg and about 30mg/kg, between about 0.1mg/kg and about 30mg/kg, between about 1mg/kg and about 30mg/kg, between about 2mg/kg and about 30mg/kg, between about 5mg/kg and about 25mg/kg, between about 5mg/kg and about 20mg/kg, between about 10mg/kg and about 20mg/kg, or between about 12mg/kg and about 18mg/kg, such as about 15mg/kg + -2 mg/kg, about 15mg/kg + -1 mg/kg, about 15mg/kg + -0.5 mg/kg, about 15mg/kg + -0.2 mg/kg, or about 15 mg/0.1 mg/kg). In some cases, the method comprises administering to the individual about 15mg/kg of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attritumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) is administered intravenously to an individual (e.g., a human) at a dose of 1200mg every three weeks (q 3 w). The dose may be administered as a single dose or as multiple doses (e.g., 2, 3, 4, 5, 6, 7, or more than 7 doses), such as infusion.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered at a dose of about 840mg every two weeks, e.g., intravenously.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered at a dose of about 1200mg every three weeks, e.g., intravenously.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered at a dose of about 1680mg every four weeks, e.g., intravenously.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered intravenously (e.g., by infusion) over 60 minutes. In some cases, for example, if the first dose is tolerated, subsequent doses may be administered intravenously (e.g., by infusion) over a 30 minute period.

In some cases, atelizumab may be administered intravenously at a dose of about 840mg every two weeks.

In some cases, atelizumab may be administered intravenously at a dose of about 1200mg every three weeks.

In some cases, the atelizumab may be administered at a dose of about 1680mg every four weeks, e.g., intravenously.

In some cases, atelizumab may be administered intravenously at a dose of 840mg every two weeks.

In some cases, atelizumab may be administered intravenously at a dose of 1200mg every three weeks.

In some cases, the atelizumab may be administered at a dose of 1680mg every four weeks, e.g., intravenously.

The atezumab can be administered intravenously (e.g., by infusion) over 60 minutes. In some cases, for example, if tolerated for the first dose, subsequent doses of atuzumab may be administered intravenously (e.g., by infusion) over a period of 30 minutes.

The dose of antibody administered in the combination therapy can be reduced compared to monotherapy. The progress of the therapy can be readily monitored by conventional techniques. In one instance, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atlizumab) is administered to the individual as a monotherapy to treat cancer. In other instances, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atlizumab) is administered to the individual as a combination therapy (as described herein) to treat cancer.

G. Indications of

The methods and medicaments described herein can be used to treat a patient having cancer (e.g., lung cancer (e.g., NSCLC), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), or breast cancer (e.g., TNBC)) by administering to the individual an effective amount of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attrituzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)). For example, the cancer can be lung cancer, kidney cancer, bladder cancer, breast cancer, colorectal cancer, ovarian cancer, pancreatic cancer, stomach cancer, esophageal cancer, mesothelioma, melanoma, head and neck cancer, thyroid cancer, sarcoma, prostate cancer, glioblastoma, cervical cancer, thymus cancer, leukemia, lymphoma, myeloma, mycosis fungoides, merkel cell carcinoma, or a hematological malignancy.

In some cases, the cancer is lung cancer. For example, the lung cancer may be non-small cell lung cancer (NSCLC), including but not limited to locally advanced or metastatic (e.g., stage IIIB, stage IV, or recurrence) NSCLC. In some cases, the lung cancer (e.g., NSCLC) is unresectable/inoperable lung cancer (e.g., NSCLC). In some cases, the lung cancer is lung cancer that has not received chemotherapy (e.g., metastatic NSCLC (mcnscc) that has not received chemotherapy). In some cases, the lung cancer is non-squamous lung cancer (e.g., non-squamous mslc). In some cases, the lung cancer is stage IV lung cancer (e.g., stage IV nsclc). In some cases, the lung cancer is recurrent lung cancer (e.g., recurrent mcnscc). In some cases, patients with lung cancer (e.g., NSCLC) have genomic alterations to EGFR or ALK. In some cases, lung cancer patients with EGFR or ALK genomic alterations develop disease progression/treatment intolerance when they receive one or more approved Tyrosine Kinase Inhibitors (TKIs).

In some cases, the cancer may be bladder cancer. For example, the bladder cancer may be Urothelial Cancer (UC), including, but not limited to, non-muscle invasive urothelial cancer, or metastatic urothelial cancer. In some cases, the urothelial cancer is metastatic urothelial cancer.

In some cases, the cancer may be renal cancer. For example, the renal cancer may be Renal Cell Carcinoma (RCC), including stage I RCC, stage II RCC, stage III RCC, stage IV RCC, or recurrent RCC.

In some cases, the cancer may be breast cancer. In some cases, the breast cancer may be a triple negative breast cancer. For example, the breast cancer can be triple negative breast cancer, estrogen receptor positive/HER 2 negative breast cancer, HER2 positive breast cancer, estrogen receptor negative breast cancer, progestin receptor positive breast cancer, or progestin receptor negative breast cancer.

In some cases, an individual having cancer (e.g., a cancer as described herein) has not been treated for the cancer. For example, an individual with cancer has not previously received a PD-L1 axis binding antagonist therapy (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as acilizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In some cases, an individual with cancer has been treated for cancer. In some cases, an individual with cancer has previously received treatment comprising a non-PD-L1 axis binding antagonist therapy (e.g., an anti-cancer therapy (e.g., a cytotoxic agent, a growth inhibitory agent, a radiation therapy, an anti-angiogenic agent, or a combination thereof)).

H. Combination therapy

In any of the methods described herein, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with an effective amount of one or more additional therapeutic agents. Suitable additional therapeutic agents include, for example, antineoplastic agents, chemotherapeutic agents, growth inhibitory agents, cytotoxic agents, radiation therapy, or combinations thereof.

In some cases, the method further comprises administering to the patient an effective amount of one or more additional therapeutic agents. In some cases, the additional therapeutic agent is selected from the group consisting of: cytotoxic agents, chemotherapeutic agents, growth inhibitory agents, radiotherapeutic agents, anti-angiogenic agents, and combinations thereof. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with a chemotherapy or chemotherapeutic agent. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with a radiotherapeutic agent. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with a targeted therapy or targeted therapeutic. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with an immunotherapy or immunotherapeutic agent (e.g., a monoclonal antibody). In some cases, the additional therapeutic agent is an agonist that targets an activating costimulatory molecule. In some cases, the additional therapeutic agent is an antagonist that targets an inhibitory co-stimulatory molecule.

Such combination therapies described above encompass both combined administration (where two or more therapeutic agents are included in the same or separate formulations) and separate administration, where administration of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) described herein can be performed before, concurrently with, and/or after administration of an additional therapeutic agent or agents. In one instance, administration of the PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) and administration of the additional therapeutic agent occur within about one month or within about one, two, or three weeks or within about one, two, three, four, five, or six days of each other.

Without wishing to be bound by theory, it is believed that enhancing T cell stimulation by promoting activation of co-stimulatory molecules or inhibiting negative co-stimulatory molecules can promote tumor cell death, thereby treating or delaying the progression of cancer. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with an agonist that targets an activating costimulatory molecule. In some cases, the activating co-stimulatory molecule may comprise CD40, CD226, CD28, OX40, GITR, CD137, CD27, HVEM, or CD127. In some cases, the agonist targeting the activating co-stimulatory molecule is an agonist antibody that binds to CD40, CD226, CD28, OX40, GITR, CD137, CD27, HVEM, or CD127. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with an antagonist that targets an inhibitory co-stimulatory molecule. In some cases, the inhibitory co-stimulatory molecule may comprise CTLA-4 (also known as CD 152), TIM-3, BTLA, VISTA, LAG-3, B7-H4, IDO, TIGIT, MICA/B, or arginase. In some cases, the antagonist that targets the inhibitory co-stimulatory molecule is an antagonist antibody that binds to CTLA-4, TIM-3, BTLA, VISTA, LAG-3, B7-H4, IDO, TIGIT, MICA/B, or arginase.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atelizumab) can be administered in combination with an antagonist (e.g., a blocking antibody) that targets CTLA-4 (also known as CD 152). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atelizumab) can be conjugated to an ipilimumab (also referred to as MDX-010, MDX-101, or

) The administration is combined. In some cases, PD-L1 axis binding antagonists(e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., alemtuzumab) can be administered in combination with tremelimumab (also known as ticilimumab or CP-675,206). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atelizumab) can be administered in combination with an antagonist (e.g., a blocking antibody) that targets B7-H3 (also referred to as CD 276). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atuzumab) may be administered in combination with MGA 271. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., altlizumab) can be administered in combination with an antagonist that targets TGF- β (e.g., metelimumab (also known as CAT-192), fresolimumab (also known as GC 1008), or LY 2157299).

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., attrituximab) can be administered in combination with a therapy comprising adoptive transfer of T cells (e.g., cytotoxic T cells or CTLs) that express a Chimeric Antigen Receptor (CAR). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atuzumab) may be administered in combination with a therapy comprising adoptive transfer of T cells comprising a dominant negative TGF β receptor (e.g., a dominant negative TGF β type II receptor). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., attentizumab) can be administered in combination with a treatment comprising a HERCREEM regimen (see, e.g., clinical trials. Gov identifier NCT 00889954).

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atlizumab) can be administered in combination with an agonist (e.g., an activating antibody) that targets CD137 (also known as TNFRSF9, 4-1BB, or ILA). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with urelumab (also referred to as BMS-663513). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with an agonist (e.g., an activating antibody) that targets CD 40. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., altlizumab) can be administered in combination with CP-870893. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with an agonist (e.g., an activating antibody) that targets OX40 (also referred to as CD 134). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., altuzumab) can be administered in combination with an anti-OX 40 antibody (e.g., agonOX). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with an agonist (e.g., an activating antibody) that targets CD 27. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with CDX-1127. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atelizumab) can be administered in combination with an antagonist that targets indoleamine-2, 3-dioxygenase (IDO). In some cases, the IDO antagonist is 1-methyl-D-tryptophan (also referred to as 1-D-MT).

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with the antibody drug conjugate. In some cases, the antibody drug conjugate comprises mertansine or monomethyl auristatin E (MMAE). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atelizumab) can be administered in combination with an anti-NaPi 2b antibody-MMAE conjugate (also referred to as DNIB0600A or RG 7599). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atuzumab) can be combined with enritumumab (also referred to as T-DM1, ado-trastuzumab emtansine or

Gene tack) in combination. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., attentizumab) can be administered in combination with DMUC 5754A. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., attrituximab) can be administered in combination with an antibody drug conjugate that targets the endothelin B receptor (EDNBR) (e.g., a conjugate of an antibody that targets EDNBR and MMAE).

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with an anti-angiogenic agent. In some cases, sub>A PD-L1 axis binding antagonist (e.g., sub>A PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atelizumab) can be administered in combination with an antibody that targets VEGF (e.g., VEGF-Sub>A). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atelizumab) can be combined with bevacizumab (also referred to as bevacizumab)

Gene tack) in combination. For example, attritumab may be administered in combination with bevacizumab. In other cases, atelizumab may be administered in combination with bevacizumab and one or more chemotherapeutic agents (e.g., carboplatin and/or paclitaxel). In certain instances, atelizumab may be administered in combination with bevacizumab, carboplatin, and paclitaxel. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., altlizumab) can be administered in combination with an antibody that targets angiopoietin 2 (also known as Ang 2). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atlizumab) can be administered in combination with MEDI 3617.

A VEGF antagonist (e.g., bevacizumab) administered to a subject (e.g., a human) in combination with a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., altlizumab) may be in the range of 0.01mg/kg to about 50mg/kg of the subject's body weight (e.g., between about 0.01mg/kg to about 45mg/kg, between about 0.01mg/kg to about 40mg/kg, between about 0.01mg/kg to about 35mg/kg, between about 0.01mg/kg to about 30mg/kg, between about 0.1mg/kg to about 30mg/kg, between about 1mg/kg to about 30mg/kg, between about 2mg/kg to about 30mg/kg, between about 5mg/kg to about 25mg/kg, between about 5mg/kg to about 20mg/kg, between about 10mg/kg to about 20mg/kg, about 15mg/kg, such as about 15mg/kg, or about 0.15 mg/kg, about 0.1mg/kg to about 15 mg/kg). For example, in some cases, the method comprises administering to the individual about 1200mg of a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attritumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) in combination with about 15mg/kg of the individual's body weight of a VEGF antagonist (e.g., bevacizumab). The method may further comprise administering one or more chemotherapeutic agents, such as carboplatin and/or paclitaxel.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with an anti-neoplastic agent. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with a drug that targets CSF-1R (also referred to as M-CSFR or CD 115). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atelizumab) can be administered in combination with an anti-CSF-1R (also referred to as IMC-CS 4). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atelizumab) can be administered in combination with an interferon (e.g., interferon alpha or interferon gamma). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., altlizumab) can be administered in combination with roscovitine-a (also known as recombinant interferon alpha-2 a). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be conjugated to GM-CSF (also Referred to as recombinant human granulocyte macrophage colony-stimulating factor, rhu GM-CSF, sargramostim or

) The administration is combined. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atelizumab) can be conjugated to IL-2 (also known as aldesleukin or alemtuzumab)

) The administration is combined. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with IL-12. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with an antibody that targets CD 20. In some cases, the CD 20-targeting antibody is obinutuzumab (also known as GA101 or GA 101)

) Or rituximab. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with an antibody that targets GITR. In some cases, the antibody targeting GITR is TRX518.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with a cancer vaccine. In some cases, the cancer vaccine is a peptide cancer vaccine, which in some cases is a personalized peptide vaccine. In some cases, the peptide Cancer vaccine is a multivalent long peptide, polypeptide, peptide mixture, hybrid peptide, or peptide-loaded dendritic cell vaccine (see, e.g., yamada et al, cancer sci.104:14-21, 2013). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with an adjuvant. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., a Tezumab) can be combined with TLR agonists (e.g., poly-ICLC (also known as

) LPS, MPL or CpG ODN). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atelizumab) can be administered in combination with Tumor Necrosis Factor (TNF) alpha (TNF-alpha). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with IL-1. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with HMGB 1. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with an IL-10 antagonist. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with an IL-4 antagonist. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with an IL-13 antagonist. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atlizumab) can be administered in combination with an HVEM antagonist. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., altuzumab) can be administered in combination with an ICOS agonist, e.g., by administering ICOS-L or an agonistic antibody that targets ICOS. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., altlizumab) can be administered in combination with a CX3CL 1-targeted therapy. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atlizumab) can be administered in combination with a treatment that targets CXCL 9. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atelizumab) can be combined with Treatment targeting CXCL10 is administered in combination. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atlizumab) may be administered in combination with a therapy that targets CCL 5. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., altlizumab) can be administered in combination with an LFA-1 or ICAM1 agonist. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with a selectin agonist.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with a targeted therapy. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with an inhibitor of B-Raf. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atelizumab) can be combined with vemurafenib (also referred to as vemurafenib)

) The administration is combined. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., attentizumab) can be combined with dabrafenib (also known as dabrafenib)

) The administration is combined. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., attentizumab) can be combined with erlotinib (also known as erlotinib)

) The administration is combined. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., attentizumab) can be administered in combination with an inhibitor of MEK, such as MEK1 (also known as MAP2K 1) or MEK2 (also known as MAP2K 2). In some cases, PD-L1 axis binding antagonists(e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., astuzumab) can be administered in combination with cobicistinib (also known as GDC-0973 or XL-518). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atelizumab) can be combined with trametinib (also referred to as tremelimumab)

) The administration is combined. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with an inhibitor of K-Ras. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with an inhibitor of c-Met. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with Onartuzumab (also known as MetMAb). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with an inhibitor of Alk. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., altuzumab) can be administered in combination with AF802 (also known as CH5424802 or aletinib). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atelizumab) can be administered in combination with a phosphatidylinositol 3-kinase (PI 3K). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atlizumab) can be administered in combination with BKM 120. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atlizumab) can be administered in combination with Idelalisib (also known as GS-1101 or CAL-101). In some embodiments, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atelizumab) can be administered in combination with pirifolin (also known as KRX-0401). In some embodiments, the PD-L1 axis binding antagonist (e.g., PD-L1 binding) An antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with an inhibitor of Akt. In some embodiments, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with MK 2206. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., attentizumab) can be administered in combination with GSK 690693. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atlizumab) can be administered in combination with GDC-0941. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., altuzumab) can be administered in combination with an inhibitor of mTOR. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atelizumab) can be administered in combination with sirolimus (also referred to as rapamycin). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atelizumab) can be combined with temsirolimus (also referred to as CCI-779 or as

) The administration is combined. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., attrituximab) can be administered in combination with everolimus (also referred to as RAD 001). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atlizumab) can be administered in combination with Ridaforolimus (also known as AP-23573, MK-8669, or Deforolimus). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atlizumab) can be administered in combination with OSI-027. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., altlizumab) can be administered in combination with AZD 8055. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with INK 128.In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with a dual PI3K/mTOR inhibitor. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., altuzumab) can be administered in combination with XL 765. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atlizumab) can be administered in combination with GDC-0980. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atelizumab) may be administered in combination with BEZ235 (also referred to as NVP-BEZ 235). In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with BGT 226. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, e.g., an anti-PD-L1 antibody, e.g., atuzumab) may be administered in combination with GSK 2126458. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atlizumab) can be administered in combination with PF-04691502. In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist, such as an anti-PD-L1 antibody, e.g., atuzumab) can be administered in combination with PF-05212384 (also known as PKI-587).

(i) Combination therapy in clinical trials

A PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attritumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered to an individual in combination with one or more additional therapeutic agents, wherein prior to or after treatment the individual has undergone diagnostic testing according to any of the diagnostic methods described herein and has been identified as an individual who may benefit from treatment with a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attritumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)). As described further below, the additional therapeutic agent may have been tested or is being tested in a clinical trial for cancer therapy including attentizumab.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attrituzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with obinutuzumab and Polatuzumab vedotin (e.g., in the treatment of lymphoma (e.g., relapsed or refractory follicular lymphoma or diffuse large B-cell lymphoma), as described in clinical trial NCT 02729896.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attrituzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be combined with paclitaxel (e.g., albumin-bound paclitaxel (nab-paclitaxel))

) Co-administration, e.g., in the treatment of breast cancer (e.g., TNBC), as described in clinical trial NCT 02530489.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atelizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be conjugated to bevacizumab (also referred to as an anti-PD-1 antibody)

) In combination (e.g. in the treatment of locally advanced or metastatic tumours (e.g. in the treatment of breast, cervical, renal, gastric, ovarian or bladder cancer) as described in clinical trial NCT 01633970.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be combined with bevacizumab (also referred to as an anti-PD-1 antibody))

) And folinic acid/oxaliplatin/5-fluorouracil (FOLFOX) (e.g., in the treatment of locally advanced or metastatic tumors, e.g., in breast cancer, cervical cancer Renal, gastric, ovarian or bladder cancer) as described in clinical trial NCT 01633970.

) And carboplatin (e.g.,

) In combination (e.g. in the treatment of locally advanced or metastatic tumours, for example in the treatment of lung cancer (NSCLC), breast cancer, cervical cancer, renal cancer, gastric cancer, ovarian cancer or bladder cancer), as described in clinical trial NCT 01633970.

) In the treatment of locally advanced or metastatic tumors (e.g. in the treatment of lung cancer (NSCLC), breast cancer, cervical cancer, renal cancer, gastric cancer, ovarian cancer or bladder cancer), as described in clinical trial NCT 01633970.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be combined with pemetrexed (e.g.,

) And carboplatin (for example,

) Combined administration (e.g. in the local evening)In the treatment of stage or metastatic tumours, for example in the treatment of breast, cervical, renal, gastric, ovarian or bladder cancer), as described in clinical trial NCT 01633970.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be conjugated to an etoposide (e.g.,

) And carboplatin (for example,

) In combination administration (e.g., in the treatment of lung cancer (e.g., small Cell Lung Cancer (SCLC)) as described in clinical trial NCT 02748889.

) And carboplatin (for example,

) In combination (e.g. in the treatment of locally advanced or metastatic tumours, for example in the treatment of lung cancer (NSCLC)), as described in clinical trial NCT 02716038.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with an Epacadostat (e.g., INCB 024360) (e.g., in the treatment of lung cancer (e.g., NSCLC) or bladder cancer (e.g., urothelial cancer)), as described in clinical trial NCT 02298153.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attrituzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with radiation therapy and chemotherapy (e.g., carboplatin and/or paclitaxel), for example, in the treatment of lung cancer (e.g., NSCLC), as described in clinical trial NCT 02525757.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with veliparib (e.g., in the treatment of breast cancer (e.g., TNBC, BRCA1 gene mutation, BRCA2 gene mutation, estrogen receptor negative breast cancer, her2/Neu negative breast cancer, stage IIIA breast cancer, stage IIIB breast cancer, stage IIIC breast cancer, or stage IV breast cancer)), as described in clinical trial NCT 02849496.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as alemtuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be conjugated to alendronide (also referred to as an anti-PD-1 antibody)

) Combined administration (e.g., in the treatment of lung cancer (e.g., NSCLC)) as described in clinical trial NCT 02013219.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be conjugated to erlotinib (also referred to as erlotinib)

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attrituximab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with MTIG7192A (e.g., in the treatment of advanced metastatic tumors), as described in clinical trial NCT 02794571.

In some cases, PD-L1 axis binding antagonismAn anti-agent (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attrituximab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be conjugated to vemurafenib (also referred to as an anti-PD-1 antibody)

) In combination (e.g. in the treatment of skin cancer (e.g. malignant melanoma)) as described in clinical trial NCT 01656642.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be combined with vemurafenib (also referred to as an anti-PD-1 antibody))

) And cobicistinib (also known as cobicistinib)

Gene tag company) in combination (e.g., in the treatment of ovarian, fallopian tube, or peritoneal carcinoma), as described in clinical trial NCT 0283907.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with obinutuzumab (e.g., in the treatment of lymphoma (e.g., lymphocytic lymphoma or relapsed refractory or Chronic Lymphocytic Leukemia (CLL)), as described in clinical trial NCT 02846623.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as altuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with carboplatin and pemetrexed (e.g., in the treatment of lung cancer (e.g., NSCLC)), as described in clinical trial NCT 02657434.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with cisplatin and pemetrexed (e.g., in the treatment of lung cancer (e.g., NSCLC)), as described in clinical trial NCT 02657434.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with Tazemetostat (e.g., in the treatment of a lymphoma (e.g., follicular lymphoma or diffuse large b-cell lymphoma)) as described in clinical trial NCT 20802242.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with obintuuzumab (e.g., in the treatment of lymphoma (e.g., follicular lymphoma or diffuse large b-cell lymphoma)), as described in clinical trial NCT 20802242.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with lenalidomide (e.g., in the treatment of multiple myeloma), as described in clinical trial NCT 02431208.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with daratuzumab (e.g., in the treatment of multiple myeloma), as described in clinical trial NCT 02431208.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with daratuzumab and lenalidomide (e.g., in the treatment of multiple myeloma), as described in clinical trial NCT 02431208.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as altuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with daratuzumab and pomalidomide (e.g., in the treatment of multiple myeloma), as described in clinical trial NCT 02431208.

Gene tack) in combination (e.g., in the treatment of renal cancer (e.g., renal cell carcinoma), as described in clinical trial NCT 02420821.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with stereotactic somatic radiation (e.g., in the treatment of lung cancer (e.g., NSCLC)), as described in clinical trial NCT 02400814.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attritumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with rociletinib (e.g., in the treatment of lung cancer (e.g., NSCLC)), as described in clinical trial NCT 02630186.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attritumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with GDC-0919 (e.g., in the treatment of a solid tumor (e.g., renal Cell Carcinoma (RCC), urothelial Cancer (UC), triple-negative breast cancer (TNBC), non-small cell lung cancer (NSCLC), melanoma, head and Neck Squamous Cell Carcinoma (HNSCC), gastric cancer, ovarian cancer, cervical cancer, endometrial cancer, or Merkel cell carcinoma), as described in clinical trial NCT 02471846.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., altuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with radium dichloride-223 (e.g., in the treatment of lung cancer, prostate cancer (e.g., castration resistant prostate cancer)), as described in clinical trial NCT 02814669.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with MOXR0916 (e.g., in the treatment of a solid tumor (e.g., a locally advanced or metastatic solid tumor)), as described in clinical trial NCT 02410512.

Gene tack) and MOXR0916 (e.g., in the treatment of solid tumors (e.g., locally advanced or metastatic solid tumors), as described in clinical trial NCT 02410512.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with azacitidine (e.g., in the treatment of a solid tumor (e.g., myelodysplastic syndrome)), as described in clinical trial NCT 02508870.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atebrine)Anti) or PD-1 binding antagonist (e.g., anti-PD-1 antibody) can be combined with paclitaxel (e.g., albumin-bound paclitaxel (nab-paclitaxel)

) Co-administration (e.g., in the treatment of breast cancer (e.g., TNBC)), as described in clinical trial NCT 02425891.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with lenalidomide and ouabituzumab (e.g., in the treatment of lymphoma), as described in clinical trial NCT 02631577.

) And carboplatin (for example,

) In combination (e.g., in the treatment of lung cancer (e.g., small Cell Lung Cancer (SCLC)), as described in clinical trial NCT 02763579.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as alemtuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with an ipilimumab (e.g., in the treatment of a locally advanced or metastatic solid tumor), as described in clinical trial NCT 02174172.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attrituximab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with interferon alpha-2 b (e.g., in the treatment of a locally advanced or metastatic solid tumor (e.g., NSCLC, melanoma, or RCC)) as described in clinical trial NCT 02174172.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with macro-fractionated image-guided radiotherapy (e.g., in the treatment of lung cancer (e.g., NSCLC)), as described in clinical trial NCT 02463994.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., astuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with CDX-1401 (e.g., in the treatment of lung cancer (e.g., NSCLC)), as described in clinical trial NCT 02495636.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attrituzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with CDX-1401 (e.g., in the treatment of lung cancer (e.g., NSCLC)), as described in clinical trial NCT 02495636.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with trastuzumab and pertuzumab (e.g., in the treatment of breast cancer (e.g., her2 positive breast cancer)), as described in clinical trial NCT 05026915.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with enrmetuzumab (e.g., in the treatment of breast cancer (e.g., her2 positive breast cancer)), as described in clinical trial NCT 02605915.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with doxorubicin and cyclophosphamide (e.g., in the treatment of breast cancer (e.g., her2 positive breast cancer)), as described in clinical trial NCT 02605915.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., altuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with trastuzumab, pertuzumab, and docetaxel (e.g., in the treatment of breast cancer (e.g., her2 positive breast cancer)), as described in clinical trial NCT 02605915.

) In combination (e.g., in the treatment of renal cancer (e.g., advanced non-clear cell renal cancer)), as described in clinical trial NCT 02724878.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with CMB305 (e.g., in the treatment of a sarcoma (e.g., mucoid/round cell liposarcoma, synovial sarcoma, metastatic sarcoma, adult recurrent soft tissue sarcoma, locally advanced sarcoma, or liposarcoma), as described in clinical trial NCT 02609984.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with RO7009789 (e.g., in the treatment of a solid cancer (e.g., locally advanced and metastatic solid tumors), as described in clinical trial NCT 02304393.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be associated with bcg (also referred to as an anti-PD-1 antibody))

) In combination (e.g., in the treatment of bladder cancer (e.g., non-muscle invasive bladder cancer)) as described in clinical trial NCT 02792192.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with stereotactic physical radiotherapy (e.g., in the treatment of lung cancer (e.g., NSCLC)), as described in clinical trial NCT 02599454.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be combined with carboplatin and albumin-bound paclitaxel (also referred to as carboplatin and albumin-bound paclitaxel)

Co-administration) co-administration (e.g., in the treatment of breast cancer (e.g., breast invasive ductal carcinoma) as described in clinical trial NCT 02620280.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be combined with carboplatin, albumin-bound paclitaxel (also known as albumin-bound paclitaxel))

) And adjuvant chemotherapy (including AC or EC (doxorubicin or epirubicin and cyclophosphamide) or FEC (fluorouracil, epirubicin and cyclophosphamide)) in combination (e.g., in the treatment of breast cancer (e.g., breast invasive ductal carcinoma), as described in clinical trial NCT 02620280.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with gemcitabine and carboplatin or cisplatin (e.g., in the treatment of urothelial cancer), as described in clinical trial NCT 02807636.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attrituximab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with paclitaxel and carboplatin (e.g., in the treatment of lung cancer (e.g., NSCLC, e.g., non-squamous NSCLC)), as described in clinical trial NCT 02366143.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with bevacizumab, paclitaxel, and carboplatin (e.g., in the treatment of lung cancer (e.g., NSCLC, e.g., non-squamous NSCLC)), as described in clinical trial NCT 02366143.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attrituzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with cerglutuzumab (also referred to as RO 6895882) (e.g., in the treatment of locally advanced and/or metastatic solid tumors), as described in clinical trial NCT 02350673.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with bendamustine and ouabituzumab (e.g., in the treatment of lymphoma (e.g., diffuse large B-cell lymphoma or follicular lymphoma), as described in clinical trial NCT 02596971.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as altuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with bendamustine, cyclophosphamide, obinutuzumab, prednisone, and vincristine (e.g., in the treatment of lymphoma (e.g., diffuse large B-cell lymphoma or follicular lymphoma), as described in clinical trial NCT 02596971.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as alemtuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with cyclophosphamide, doxorubicin, obinutuzumab, prednisone, and vincristine (e.g., in the treatment of lymphoma (e.g., diffuse large B-cell lymphoma or follicular lymphoma), as described in clinical trial NCT 02596971.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attrituximab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with cyclophosphamide, doxorubicin, prednisone, vincristine and rituximab (e.g., in the treatment of lymphoma (e.g., diffuse large B-cell lymphoma or follicular lymphoma), as described in clinical trial NCT 02596971.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as alemtuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with RO6958688 (e.g., in the treatment of locally advanced and/or metastatic solid tumors (e.g., carcinoembryonic antigen (CEA) -positive solid tumors)), as described in clinical trial NCT 02650713.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with acetylsalicylic acid (e.g., in the treatment of ovarian cancer (e.g., ovarian tumor)), as described in clinical trial NCT 02659384.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with bevacizumab (e.g., in the treatment of ovarian cancer (e.g., ovarian tumor)), as described in clinical trial NCT 02659384.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attrituzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with vanucizumab (also known as RO 5520985) (e.g., in the treatment of locally advanced and/or metastatic solid tumors), as described in clinical trial NCT 01688206.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with carboplatin and an albumin-bound paclitaxel (e.g., in the treatment of lung cancer (e.g., non-squamous NSCLC), as described in clinical trial NCT 02367781.

) In combination (e.g., in the treatment of renal cancer (e.g., renal cell carcinoma)), as described in clinical trial NCT 01984242.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attentizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with cobicistinib (also referred to as GDC-0973) (e.g., in the treatment of locally advanced or metastatic solid tumors), as described in clinical trial NCT 01988896.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with RO5509554 (e.g., in the treatment of locally advanced solid tumors (e.g., locally advanced and/or metastatic triple negative breast cancer, ovarian cancer, bladder cancer, gastric cancer, or soft tissue sarcoma), as described in clinical trial NCT 02323191.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attrituzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with vallizumab (varlumab) (e.g., in the treatment of advanced cancer (e.g., melanoma, RCC, triple negative breast cancer, bladder cancer, head and neck cancer, or non-small cell lung cancer)), as used in clinical trial NCT 02543645.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attrituximab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with cobitinib (e.g., in the treatment of colorectal cancer), as described in clinical trial NCT 02788279.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with cobicistinib (e.g., in the treatment of colorectal cancer), as described in clinical trial NCT 02788279.

) Co-administration (e.g. in the treatment of solid tumours) as described in clinical trial NCT 02715531.

) Folinic acid, oxaliplatin and optionally capecitabine (e.g. in the treatment of solid tumours), as described in clinical trial NCT 02715531.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attrituximab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with albumin-bound paclitaxel and gemcitabine (e.g., in the treatment of a solid tumor), as described in clinical trial NCT 02715531.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with oxaliplatin, leucovorin, 5-fluorouracil (5-FU), oxaliplatin and cisplatin (e.g., in the treatment of a solid tumor), as described in clinical trial NCT 02715531.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as alemtuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with albumin-bound paclitaxel and carboplatin (e.g., in the treatment of lung cancer (e.g., squamous NSCLC), as described in clinical trial NCT 02367794.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attrituximab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with paclitaxel and carboplatin (e.g., in the treatment of lung cancer (e.g., squamous NSCLC), as described in clinical trial NCT 02367794.

In some cases, a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as alemtuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) can be administered in combination with CPI-444 (e.g., in the treatment of advanced cancers (e.g., non-small cell lung cancer, malignant melanoma, renal cell carcinoma, triple negative breast cancer, microsatellite instability colorectal cancer (MSI), and bladder cancer)) as described in clinical trial NCT 02655822.

PD-L1 axis binding antagonists

PD-L1 axis binding antagonists include PD-L1 binding antagonists, PD-1 binding antagonists, and PD-L2 binding antagonists. PD-1 (programmed death 1) is also known in the art as "programmed cell death 1", "PDCD1", "CD279" and "SLEB2". An exemplary human PD-L1 is shown in UniProtKB/Swiss-Prot accession number Q9NZQ7.1. Exemplary human PD-1 is shown in UniProtKB/Swiss-Prot accession number Q15116. PD-L1 (programmed death ligand 1) is also known in the art as "programmed cell death 1 ligand 1", "PDCD1LG1", "CD274", "B7-H", and "PDL1". PD-L2 (programmed death ligand 2) is also known in the art as "programmed cell death 1 ligand 2", "PDCD1LG2", "CD273", "B7-DC", "Btdc" and "PDL2". Exemplary human PD-L2 is shown in UniProtKB/Swiss-Prot accession number Q9BQ 51. In some embodiments, PD-L1, PD-1, and PD-L2 are human PD-L1, PD-1, and PD-L2. In some cases, the PD-L1 axis binding antagonist can be a PD-L1 binding antagonist, a PD-1 binding antagonist, or a PD-L2 binding antagonist.

Any of the methods, compositions for use, uses, kits or articles of manufacture described herein can include or relate to any of the PDL-L1 axis binding antagonists described below.

In one aspect, the disclosure provides PD-L1 axis binding antagonists for use in, e.g., any one of the methods disclosed herein. In one embodiment, the PD-L1 axis binding antagonist is used to treat cancer. In one such embodiment, the method further comprises administering to the individual an effective amount of at least one additional therapeutic agent, such as any of the additional therapeutic agents disclosed herein.

In another aspect, the present disclosure provides the use of a PD-L1 axis binding antagonist for the manufacture or preparation of a medicament for use in any of the methods, e.g., disclosed herein. In one embodiment, the medicament is for treating cancer. In one such embodiment, the method further comprises administering to the individual an effective amount of at least one additional therapeutic agent, such as any of the additional therapeutic agents disclosed herein.

PD-L1 binding antagonists

In some cases, the PD-L1 binding antagonist inhibits the binding of PD-L1 to one or more of its ligand binding partners. In other cases, the PD-L1 binding antagonist inhibits the binding of PD-L1 to PD-1. In still other cases, the PD-L1 binding antagonist inhibits the binding of PD-L1 to B7-1. In some cases, the PD-L1 binding antagonist inhibits the binding of PD-L1 to both PD-1 and B7-1. In some cases, the PD-L1 binding antagonist is an antibody. In some cases, the antibody is selected from the group consisting of alemtuzumab, yw243.55.S70, MDX-1105, MEDI4736 (devaluzumab), and MSB0010718C (avizumab).

In some cases, the anti-PD-L1 antibody is a monoclonal antibody. In some cases, the anti-PD-L1 antibody is selected from the group consisting of Fab, fab '-SH, fv, scFv, and (Fab') ₂ Antibody fragments of the group consisting of fragments. In some cases, the anti-PD-L1 antibody is a humanized antibody. In some cases, the anti-PD-L1 antibody is a human antibody. In some cases, an anti-PD-L1 antibody described herein binds to human PD-L1. In some particular cases, the anti-PD-L1 antibody is atelizumab (CAS registry number: 1422185-06-5). Astuzumab (genethak) is also known as MPDL3280A.

In some cases, an anti-PD-L1 antibody comprises a heavy chain variable region (HVR-H) comprising HVR-H1, HVR-H2, and HVR-H3 sequences, wherein:

(a) The HVR-H1 sequence is GFTFSDSWIH (SEQ ID NO: 1);

(b) The HVR-H2 sequence is AWISPYGGSTYYADSVKG (SEQ ID NO: 2); and is

(c) The HVR-H3 sequence is RHWGGFDY (SEQ ID NO: 3).

In some cases, the anti-PD-L1 antibody further comprises a light chain variable region (HVR-L) comprising HVR-L1, HVR-L2, and HVR-L3 sequences, wherein:

(a) The HVR-L1 sequence is RASQDVSTAVA (SEQ ID NO: 4);

(b) The HVR-L2 sequence is SASFLYS (SEQ ID NO: 5); and is

(c) The HVR-L3 sequence was QQYLYHPAT (SEQ ID NO: 6).

In some cases, the anti-PD-L1 antibody comprises heavy and light chain sequences, wherein:

(a) The heavy chain Variable (VH) region sequence comprises the amino acid sequence:

<xnotran> EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSS (SEQ ID NO: 7); </xnotran> And is

(b) The light chain Variable (VL) region sequence comprises the amino acid sequence:

DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKR(SEQ ID NO:8)。

(a) The heavy chain comprises the following amino acid sequence:

<xnotran> EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 9); </xnotran> And is

(b) The light chain comprises the following amino acid sequence:

DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO:10)。

in some cases, an anti-PD-L1 antibody comprises (a) a VH domain comprising an amino acid sequence having at least 95% sequence identity (e.g., at least 95%, 96%, 97%, 98%, or 99% sequence identity) to the sequence of SEQ ID No. 7, or a sequence comprising SEQ ID No. 7; (b) A VL domain comprising an amino acid sequence having at least 95% sequence identity (e.g., at least 95%, 96%, 97%, 98%, or 99% sequence identity) to the sequence of SEQ ID No. 8, or comprising the sequence of SEQ ID No. 8; or (c) a VH domain as described in (a) and a VL domain as described in (b). In other cases, the anti-PD-L1 antibody is selected from the group consisting of: YW243.55.S70, MDX-1105, MEDI4736 (Dewaruzumab) and MSB0010718C (Avermectin). Antibody YW243.55.S70 is anti-PD-L1 as described in PCT publication No. WO 2010/077634. MDX-1105 is also known as BMS-936559, an anti-PD-L1 antibody described in PCT publication No. WO 2007/005874. MEDI4736 (Devolumab) is an anti-PD-L1 monoclonal antibody described in PCT publication No. WO 2011/066389 and U.S. patent publication No. 2013/034559. anti-PD-L1 antibodies useful in the methods of the invention and methods of making the same are described in PCT publication nos. WO 2010/077634, WO2007/005874, and WO 2011/066389, as well as U.S. patent No. 8,217,149 and U.S. patent publication No. 2013/034559, which are incorporated herein by reference.

It is expressly contemplated that such PD-L1 binding antagonist antibodies for any one of the above-listed cases can have any of the features described in sections 1-7 below, alone or in combination.

PD-1 binding antagonists

In some cases, the PD-L1 axis binding antagonist is a PD-1 binding antagonist. For example, in some cases, a PD-1 binding antagonist inhibits the binding of PD-1 to one or more of its ligand binding partners. In some cases, the PD-1 binding antagonist inhibits the binding of PD-1 to PD-L1. In other cases, the PD-1 binding antagonist inhibits the binding of PD-1 to PD-L2. In still other cases, the PD-1 binding antagonist inhibits the binding of PD-1 to both PD-L1 and PD-L2. In some cases, the PD-1 binding antagonist is an antibody. In some cases, the antibody is selected from the group consisting of: MDX 1106 (nivolumab), MK-3475 (pembrolizumab), MEDI-0680 (AMP-514), PDR001, REGN2810, and BGB-108. In some cases, the PD-1 binding antagonist is an Fc fusion protein. For example, in some cases, the Fc fusion protein is AMP-224.

In some embodiments, the PD-1 binding antagonist is a molecule that inhibits the binding of PD-1 to its ligand binding partner. In particular aspects, the PD-1 ligand binding partner is PD-L1 and/or PD-L2. In another embodiment, the PD-L1 binding antagonist is a molecule that inhibits the binding of PD-L1 to its binding partner. In particular aspects, the PD-L1 binding partner is PD-1 and/or B7-1. In another embodiment, the PD-L2 binding antagonist is a molecule that inhibits the binding of PD-L2 to its ligand binding partner. In a specific aspect, the PD-L2 binding ligand partner is PD-1. The antagonist can be an antibody, an antigen-binding fragment thereof, an immunoadhesin, a fusion protein or an oligopeptide.

In some embodiments, the PD-1 binding antagonist is an anti-PD-1 antibody (e.g., a human antibody, a humanized antibody, or a chimeric antibody), e.g., as described below. In some embodiments, the anti-PD-1 antibody is selected from the group consisting of: MDX-1106 (nivolumab), MK-3475 (pembrolizumab), MEDI-0680 (AMP-514), PDR001, REGN2810, and BGB-108.MDX-1106, also known as MDX-1106-04, ONO-4538, BMS-936558 or nivolumab, is an anti-PD-1 antibody described in WO 2006/121168. MK-3475, also known as pembrolizumab or Lambolizumab, is an anti-PD-1 antibody described in WO 2009/114335. In some embodiments, the PD-1 binding antagonist is an immunoadhesin (e.g., an immunoadhesin comprising an extracellular or PD-1 binding portion of PD-L1 or PD-L2 fused to a constant region (e.g., the Fc region of an immunoglobulin sequence)). In some embodiments, the PD-1 binding antagonist is AMP-224.AMP-224, also known as B7-DCIg, is a PD-L2-Fc fusion soluble receptor described in WO 2010/027827 and WO 2011/066342.

In some embodiments, the anti-PD-1 antibody is MDX-1106. Alternative names for "MDX-1106" include MDX-1106-04, ONO-4538, BMS-936558, and nivolumab. In some embodiments, the anti-PD-1 antibody is nivolumab (CAS registry number: 946414-94-4). In yet another embodiment, an isolated anti-PD-1 antibody is provided, which antibody comprises: a heavy chain variable region comprising the heavy chain variable region amino acid sequence of SEQ ID NO 11; and/or a light chain variable region comprising the light chain variable region amino acid sequence of SEQ ID NO 12.

In yet another embodiment, an isolated anti-PD-1 antibody is provided, which antibody comprises heavy and/or light chain sequences, wherein:

(a) The heavy chain sequence has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity to the heavy chain sequence of seq id no:

<xnotran> QVQLVESGGGVVQPGRSLRLDCKASGITFSNSGMHWVRQAPGKGLEWVAVIWYDGSKRYYADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATNDDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 11), </xnotran>

(b) The light chain sequence has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a light chain sequence that is:

EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQSSNWPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC(SEQ ID NO:12)。

It is specifically contemplated that such PD-1 binding antagonist antibodies for any one of the above-listed circumstances can have any of the features described in sections 1-7, below, alone or in combination.

C. Antibodies

1. Affinity of antibody

In certain instances, an antibody provided herein (e.g., an anti-PD-L1 antibody or an anti-PD-1 antibody) has a dissociation constant (Kd) of less than or equal to 1 μ M, less than or equal to 100nM, less than or equal to 10nM, less than or equal to 1nM, less than or equal to 0.1nM, less than or equal to 0.01nM, or less than or equal to 0.001nM (e.g., 10 nM) ^-8 M or less, e.g. 10 ^-8 M to 10 ^-13 M, e.g. 10 ^-9 M to 10 ^-13 M)。

In one instance, kd is measured by a radiolabeled antigen binding assay (RIA). In one example, RIA is performed using Fab forms of the antibody of interest and its antigen. For example, by using a minimum concentration in the presence of a series of unlabeled antigen titrations ( ¹²⁵ I) The solution binding affinity of Fab to antigen was measured by equilibration of the Fab with labeled antigen and subsequent capture of the bound antigen with an anti-Fab antibody coated plate (see, e.g., chen et al, J.mol.biol.293:865-881 (1999)). To determine the conditions for the assay, capture anti-Fab antibodies (Cappel Labs) were coated with 5. Mu.g/ml in 50mM sodium carbonate (pH 9.6)

The plate (Thermo Scientific) was blocked overnight, then for two to five hours at room temperature (about 23 ℃) with 2% (w/v) bovine serum albumin in PBS. In the non-adsorption plate (Nunc # 269620), mixing 100pM or 26pM ¹²⁵ I]Mixing of antigen with serial dilutions of Fab of interest (e.g.following the assessment of anti-VEGF antibodies (Fab-12) in Presta et al, cancer Res.57:4593-4599 (1997)). Then incubating the target Fab overnight; however, incubation may be continued for a longer period of time (e.g., about 65 hours) to ensure equilibrium is reached. Thereafter, the mixture is transferred to a capture plate for incubation at room temperature (e.g., one hour). The solution was then removed and used with 0.1% polysorbate 20 in PBS

The plate was washed eight times. When the plates had dried, 150. Mu.l/well of scintillator (MICROSCINT-20) was added ^TM (ii) a Packard) and in TOPCOUNT ^TM The gamma counter (Packard) counts the plate for tens of minutes. The concentration of each Fab that gave less than or equal to 20% maximal binding was selected for use in a competitive binding assay.

According to another case, use

Surface plasmon resonance assay measures Kd. For example, use

-2000 or

-3000 (BIAcore, inc., piscataway, NJ) was assayed at 25 ℃ with an immobilized antigen CM5 chip at about 10 Response Units (RU). In one case, carboxymethylated dextran biosensor chips (CM 5, BIACORE, inc.) were activated with N-ethyl-N '- (3-dimethylaminopropyl) -carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) according to supplier's instructions. Antigen was diluted to 5. Mu.g/ml (about 0.2. Mu.M) with 10mM sodium acetate, pH 4.8, followed by injection at a flow rate of 5. Mu.L/min to obtain approximately 10 response units A conjugated protein at position (RU). After injection of the antigen, 1M ethanolamine was injected to block unreacted groups. For kinetic measurements, injection containing 0.05% polysorbate 20 (TWEEN 20) was performed at 25 ℃ at a flow rate of about 25. Mu.L/min ^TM ) Two-fold serial dilutions (0.78 nM to 500 nM) of Fab in PBS of surfactant (PBST). Using a simple one-to-one Langmuir binding model: (

Evaluation Software version 3.2) for calculating association rates (k) by simultaneous fitting of association and dissociation sensor maps _on ) And dissociation rate (k) _off ). The equilibrium dissociation constant (Kd) is calculated as the ratio k _off /k _on . See, e.g., chen et al, J.mol.biol.293:865-881 (1999). If the association rate exceeds 10 as determined by the above surface plasmon resonance ⁶ M ^-1 s ^-1 The rate of association can then be determined by using a fluorescence quenching technique, e.g., in a spectrometer such as an Aviv Instruments or 8000 series SLM-AMINCO ^TM The increase or decrease in fluorescence emission intensity (excitation =295nM; emission =340nm, band pass at 1691m) of 20nM anti-antigen antibody (Fab form) in PBS pH 7.2 at 25 ℃ was measured in a spectrophotometer (ThermoSpectronic) with a stirred cuvette in the presence of increasing concentrations of antigen.

2. Antibody fragments

In certain instances, an antibody provided herein (e.g., an anti-PD-L1 antibody or an anti-PD-1 antibody) is an antibody fragment. Antibody fragments include, but are not limited to, fab '-SH, F (ab') ₂ Fv, and scFv fragments, as well as other fragments described below. For a review of certain antibody fragments, see Hudson et al, nat. Med.9:129-134 (2003). For reviews on scFv fragments see, for example, pluckth ü n in The pharmacogolology of Monoclonal Antibodies, vol.113, rosenburg and Moore eds. (Springer-Verlag, new York), pp.269-315 (1994); see also WO 93/16185; and U.S. Pat. nos. 5,571,894 and 5,587,458. For Fa containing salvage receptor binding epitope residues and having increased half-life in vivob and F (ab') ₂ See U.S. Pat. No. 5,869,046 for a discussion of fragments.

Diabodies are antibody fragments with two antigen binding sites, which may be bivalent or bispecific. See, for example, EP 404,097; WO 1993/01161; hudson et al, nat. Med.9:129-134 (2003); and Hollinger et al, proc.natl.acad.sci.usa 90. Tri-and tetrad antibodies are also described in Hudson et al, nat. Med.9:129-134 (2003).

A single domain antibody is an antibody fragment comprising all or part of a heavy chain variable domain or all or part of a light chain variable domain of an antibody. In certain instances, the single domain antibody is a human single domain antibody (Domantis, inc., waltham, MA; see, e.g., U.S. Pat. No. 6,248,516B1).

Antibody fragments can be prepared by a variety of techniques, including but not limited to proteolytic digestion of intact antibodies and production by recombinant host cells (e.g., e.coli or phage), as described herein.

3. Chimeric and humanized antibodies

In certain instances, an antibody provided herein (e.g., an anti-PD-L1 antibody or an anti-PD-1 antibody) is a chimeric antibody. Certain chimeric antibodies are described, for example, in U.S. Pat. No. 4,816,567 and Morrison et al, proc. Natl. Acad. Sci. USA, 81. In one example, a chimeric antibody comprises a non-human variable region (e.g., a variable region derived from a mouse, rat, hamster, rabbit, or non-human primate (such as a monkey)) and a human constant region. In another example, a chimeric antibody is a "class switch" antibody in which the class or subclass has been altered from that of the parent antibody. Chimeric antibodies include antigen-binding fragments thereof.

In certain instances, the chimeric antibody is a humanized antibody. Typically, non-human antibodies are humanized to reduce immunogenicity to humans, while retaining the specificity and affinity of the parent non-human antibody. Typically, a humanized antibody comprises one or more variable domains in which HVRs, e.g., CDRs (or portions thereof), are derived from a non-human antibody and FRs (or portions thereof) are derived from a human antibody sequence. The humanized antibody optionally will also comprise at least a portion of a human constant region. In some examples, some FR residues in a humanized antibody are replaced with corresponding residues from a non-human antibody (e.g., the antibody from which the HVR residues are derived), e.g., to restore or improve antibody specificity or affinity.

Humanized antibodies and methods for their preparation are reviewed, for example, in Almagro and Fransson, front.biosci.13:1619-1633 (2008), and are further described, for example, in Riechmann et al, nature 332; queen et al, proc. Natl. Acad. Sci. USA86:10029-10033 (1989); U.S. Pat. nos. 5,821,337, 7,527,791, 6,982,321, and 7,087,409; kashmiri et al, methods 36 (2005) (describes Specificity Determining Region (SDR) grafting); padlan, mol.Immunol.28:489-498 (1991) (described as "surface remodeling"); dall' Acqua et al, methods 36 (2005) (describing "FR shuffling"); and Osbourn et al, methods 36 (2005) and Klimka et al, br.J. cancer,83, 252-260 (2000) (describing the "guided selection" method for FR shuffling).

Human framework regions that may be used for humanization include, but are not limited to: framework regions selected using a "best fit" approach (see, e.g., sims et al J. Immunol.151:2296 (1993)); the framework regions derived from the consensus sequence of a human antibody from a particular subgroup of light or heavy chain variable regions (see, e.g., carter et al Proc. Natl. Acad. Sci. USA,89 4285 (1992); and Presta et al J. Immunol.,151 (1993)); human mature (somatic mutation) framework regions or human germline framework regions (see, e.g., almagro and Fransson, front. Biosci.13:1619-1633 (2008)); and framework regions derived from screening FR libraries (see, e.g., baca et al, J.biol. Chem.272:10678-10684 (1997) and Rosok et al, J.biol. Chem.271:22611-22618 (1996)).

4. Human antibodies

In certain instances, an antibody provided herein (e.g., an anti-PD-L1 antibody or an anti-PD-1 antibody) is a human antibody. Human antibodies can be produced using various techniques known in the art. Human antibodies are generally described in van Dijk and van de Winkel, curr Opin pharmacol.5:368-74 (2001) and Lonberg, curr Opin immunol.20:450-459 (2008).

Can pass throughHuman antibodies were prepared in the following manner: the immunogen is administered to a transgenic animal that has been modified to produce a fully human antibody or a fully antibody with human variable regions in response to antigen challenge. Such animals typically contain all or part of a human immunoglobulin locus that replaces an endogenous immunoglobulin locus, or is present extrachromosomally or randomly integrated into the chromosome of the animal. In such transgenic mice, the endogenous immunoglobulin loci have typically been inactivated. For an overview of the method for obtaining human antibodies from transgenic animals, see Lonberg, nat. Biotech.23:1117-1125 (2005). See also, for example, the description of XENOMOUSE ^TM U.S. Pat. nos. 6,075,181 and 6,150,584 to technology; description of the invention

U.S. Pat. nos. 5,770,429; description of K-M

U.S. Pat. No. 7,041,870 to the Art, and description

U.S. patent application publication No. US 2007/0061900 of the art. The human variable regions from intact antibodies produced by such animals may be further modified, for example by combination with different human constant regions.

Human antibodies can also be prepared by hybridoma-based methods. Human myeloma and mouse-human hybrid myeloma cell lines have been described for the production of human monoclonal antibodies. (see, e.g., kozbor J.Immunol.,133 (1984); brodeur et al, monoclonal Antibody Production Techniques and Applications, pp 51-63 (Marcel Dekker, inc., new York, 1987); and Boerner et al, J.Immunol.,147 (1991)), human antibodies produced via human B-cell hybridoma technology are also described by Li et al, proc.Natl.Acad.Sci.USA, 103. Additional methods include, for example, those described in U.S. Pat. No. 7,189,826 (describing the production of monoclonal human IgM antibodies from hybridoma cell lines) and Ni, xiandai Mianyixue,26 (4): 265-268 (2006) (describing human-human hybridomas). The human hybridoma technique (Trioma technique) is also described in Vollmers and Brandlein, histology and Histopathology,20 (3): 927-937 (2005) and Vollmers and Brandlein, methods and Findings in Experimental and Clinical Pharmacology,27 (3): 185-91 (2005).

Human antibodies can also be produced by isolating Fv clone variable domain sequences selected from a human phage display library. Such variable domain sequences can then be combined with the desired human constant domains. Techniques for selecting human antibodies from antibody libraries are described below.

5. Antibodies derived from libraries

Antibodies (e.g., anti-PD-L1 antibodies and anti-PD-1 antibodies) can be isolated by screening combinatorial libraries for antibodies having a desired activity. For example, various methods are known in the art for generating phage display libraries and screening such libraries for antibodies with desired binding characteristics. Such Methods are reviewed, for example, in Hoogenboom et al, methods in Molecular Biology 178, 1-37 (O' Brien et al, eds., human Press, totowa, NJ, 2001) and are further described, for example, in McCafferty et al, nature 348; clackson et al, nature 352; marks et al, J.mol.biol.222:581-597 (1992); marks and Bradbury, in Methods in Molecular Biology 248 161-175 (Lo, ed., human Press, totowa, NJ, 2003); sidhu et al, J.mol.biol.338 (2): 299-310 (2004); lee et al, J.mol.biol.340 (5): 1073-1093 (2004); fellouse, proc.natl.acad.sci.usa 101 (34); 12467-12472 (2004); and Lee et al, J.Immunol.methods 284 (1-2): 119-132 (2004).

In certain phage display methods, repertoires of VH and VL genes are individually cloned by Polymerase Chain Reaction (PCR) and randomly recombined in a phage library, from which antigen-binding phage can then be screened for, as described in Winter et al, ann.rev.immunol.,12 (1994). Phage typically display antibody fragments as single chain Fv (scFv) fragments or Fab fragments. Libraries from immunized sources provide high affinity antibodies to the immunogen without the need to construct hybridomas. Alternatively, all natural components (e.g., all natural components from humans) can be cloned to provide a single source of antibodies to a wide range of non-self and self-antigens without any immunization, as described by Griffiths et al, EMBO J, 12. Finally, the initial library can also be made by: cloning unrearranged V gene segments from stem cells; and the use of PCR primers containing randomized sequences to encode highly variable CDR3 regions and to accomplish in vitro rearrangement as described by Hoogenboom and Winter, j.mol.biol., 227. Patent publications describing human antibody phage libraries include, for example: U.S. Pat. No. 5,750,373, and U.S. publication Nos. 2005/0079574, 2005/0119455, 2005/0266000, 2007/0117126, 2007/0160598, 2007/0237764, 2007/0292936, and 2009/0002360.

Antibodies or antibody fragments isolated from a human antibody library are considered herein to be human antibodies or human antibody fragments.

6. Multispecific antibodies

In any of the above aspects, an antibody provided herein (e.g., an anti-PD-L1 antibody or an anti-PD-1 antibody) can be a multispecific antibody, e.g., a bispecific antibody. Multispecific antibodies are monoclonal antibodies having binding specificities for at least two different sites. In certain instances, the antibodies provided herein are multispecific antibodies, e.g., bispecific antibodies. In certain instances, one of the binding specificities is for PD-L1 and the other is for any other antigen. In certain instances, a bispecific antibody can bind two different epitopes of PD-L1. Bispecific antibodies may also be used to localize cytotoxic agents to cells expressing PD-L1. Bispecific antibodies can be prepared as full length antibodies or antibody fragments.

Techniques for making multispecific antibodies include, but are not limited to, recombinant co-expression of two immunoglobulin heavy chain-light chain pairs with different specificities (see, milstein and Cuello, nature 305 (1983), WO 93/08829 and Traunecker et al, EMBO j.10:3655 (1991)) and "hole-and-mortar" engineering (see, e.g., U.S. Pat. No. 5,731,168). Multispecific antibodies can also be made by the following techniques: engineering electrostatically manipulated effects to make antibody Fc-heterodimer molecules (see, e.g., WO 2009/089004 A1); cross-linking two or more antibodies or fragments (see, e.g., U.S. Pat. No. 4,676,980 and Brennan et al, science 229 (1985)); the use of leucine zippers to generate bispecific antibodies (see, e.g., kostelny et al, J.Immunol.148 (5): 1547-1553 (1992)); bispecific antibody fragments were made using the "diabody" technique (see, e.g., hollinger et al, proc. Natl. Acad. Sci. USA90:6444-6448 (1993)); single chain Fv (sFv) dimers were used (see, e.g., gruber et al, J.Immunol.152:5368 (1994)); and trispecific antibodies were prepared as described, for example, in Tutt et al J.Immunol.147:60 (1991).

Also included herein are engineered antibodies having three or more functional antigen binding sites, including "octopus antibodies" (see, e.g., US 2006/0025576 A1).

The antibodies or fragments herein also include "dual action fabs" or "DAFs" that comprise an antigen binding site that binds to PD-L1 as well as another, different antigen.

7. Antibody variants

In certain instances, amino acid sequence variants of the antibodies provided herein (e.g., anti-PD-L1 antibodies and anti-PD-1 antibodies) are contemplated. For example, it may be desirable to improve the binding affinity and/or other biological properties of an antibody. Amino acid sequence variants of an antibody can be prepared by introducing appropriate modifications into the nucleotide sequence encoding the antibody or by peptide synthesis. Such modifications include, for example, deletions from, and/or insertions into, and/or substitutions of, residues within the amino acid sequence of the antibody. Any combination of deletions, insertions, and substitutions can be made to arrive at the final construct, provided that the final construct possesses the desired characteristics, e.g., antigen binding.

I. Substitution, insertion and deletion variants

In certain instances, antibody variants are provided having one or more amino acid substitutions. Sites of interest for substitution mutations include HVRs and FRs. Conservative substitutions are shown in table a under the heading of "preferred substitutions". Further substantial changes are provided under the heading "exemplary substitutions" in table a, and are further described below with reference to amino acid side chain classes. Amino acid substitutions can be introduced into the antibody of interest and the product screened for a desired activity (e.g., retained/improved antigen binding, reduced immunogenicity, or improved antibody-dependent cell-mediated cytotoxicity (ADCC) or complement-dependent cytotoxicity (CDC)).

Exemplary and preferred amino acid substitutions

Amino acids can be grouped according to common side chain properties:

(1) Hydrophobicity: norleucine, met, ala, val, leu, ile;

(2) Neutral hydrophilicity: cys, ser, thr, asn, gln;

(3) Acidity: asp and Glu;

(4) Alkalinity: his, lys, arg;

(5) Residues that influence chain orientation: gly, pro;

(6) Aromatic: trp, tyr, phe.

Non-conservative substitutions will require the exchange of a member of one of these classes for another.

One type of substitution variant involves substituting one or more hypervariable region residues of a parent antibody (e.g., a humanized or human antibody). Typically, one or more of the resulting variants selected for further study will have modifications (e.g., improvements) in certain biological properties (e.g., increased affinity and/or reduced immunogenicity) relative to the parent antibody and/or will have certain biological properties of the parent antibody that are substantially retained. Exemplary substitution variants are affinity matured antibodies, which can be conveniently generated using phage display-based affinity maturation techniques such as those described herein, for example. Briefly, one or more HVR residues are mutated and variant antibodies are displayed on phage and screened for a particular biological activity (e.g., binding affinity).

For example, HVRs can be altered (e.g., substituted) to improve antibody affinity. Such changes can occur in HVR "hotspots", i.e., residues encoded by codons that undergo high frequency mutations during somatic maturation (see, e.g., chowdhury, methods mol. Biol.207:179-196 (2008)) and/or residues that come into contact with antigen (detecting binding affinity of the resulting variant VH or VL. Affinity maturation achieved by construction and reselection from secondary libraries has been described, for example, by Hoogenboom et al in Methods in Molecular Biology 178 (O' Brien et al, human Press, totowa, NJ, (2001)).

In certain examples, substitutions, insertions, or deletions may occur within one or more HVRs, so long as such alterations do not substantially reduce the ability of the antibody to bind antigen. For example, conservative changes that do not substantially reduce binding affinity (e.g., conservative substitutions as provided herein) may be made in HVRs. Such changes may be, for example, outside of antigen-contacting residues in HVRs. In certain examples of the variant VH and VL sequences provided above, each HVR is unchanged, or contains no more than one, two, or three amino acid substitutions.

A method that can be used to identify antibody residues or regions that can be targeted for mutagenesis is referred to as "alanine scanning mutagenesis" as described by Cunningham and Wells (1989) Science,244 1081-1085. In this method, a residue or set of target residues (e.g., charged residues such as Arg, asp, his, lys, and Glu) are identified and replaced with a neutral or negatively charged amino acid (e.g., alanine or polyalanine) to determine whether antibody interaction with an antigen is affected. Additional substitutions may be introduced at amino acid positions that exhibit functional sensitivity to the initial substitution. Alternatively or additionally, the crystal structure of the antigen-antibody complex is used to identify contact points between the antibody and the antigen. Such contact residues and adjacent residues that are candidates for substitution may be targeted or eliminated. Variants can be screened to determine if they possess the desired properties.

Amino acid sequence insertions include amino and/or carboxyl terminal fusions ranging in length from one residue to polypeptides containing a hundred or more residues, as well as intrasequence insertions of one or more amino acid residues. Examples of terminal insertions include antibodies with an N-terminal methionyl residue. Other insertional variants of the antibody molecule include fusions to the N-terminus or C-terminus of an antibody of an enzyme (e.g., for ADEPT) or polypeptide that increases the serum half-life of the antibody.

Glycosylation variants

In certain instances, the antibodies of the invention can be altered to increase or decrease the extent to which the antibody is glycosylated. The addition or deletion of glycosylation sites of the antibodies of the invention can be conveniently achieved by altering the amino acid sequence to create or remove one or more glycosylation sites.

When the antibody comprises an Fc region, the carbohydrate attached thereto may be altered. Natural antibodies produced by mammalian cells typically comprise branched, bi-antennary oligosaccharides typically linked by an N-bond to Asn297 of the CH2 domain of the Fc region. See, e.g., wright et al TIBTECH 15 (1997). Oligosaccharides may include various carbohydrates, for example, mannose, N-acetylglucosamine (GlcNAc), galactose, and sialic acid, as well as fucose attached to GlcNAc in the "backbone" of the biantennary oligosaccharide structure. In some examples, the oligosaccharides in the antibodies of the invention may be modified to produce antibody variants with certain improved properties.

In one instance, antibody variants are provided that have a carbohydrate structure that lacks fucose attached (directly or indirectly) to an Fc region. For example, the fucose content in such antibodies may be 1% to 80%, 1% to 65%, 5% to 65%, or 20% to 40%. The amount of fucose is determined by calculating the average amount of fucose at Asn297 in the sugar chain, relative to the sum of all sugar structures (e.g., complex, hybrid, and high mannose structures) attached to Asn297 as determined by MALDI-TOF mass spectrometry, as described in WO 2008/077546. Asn297 refers to the asparagine residue at about position 297 in the Fc region (EU numbering of Fc region residues); however, due to minor sequence variations in antibodies, asn297 may also be located approximately ± 3 amino acids upstream or downstream of position 297, i.e. between

positions

294 and 300. Such fucosylated variants may have improved ADCC function. See, for example, U.S. patent publication Nos. US 2003/0157108 and US 2004/0093621. Examples of publications relating to "defucosylated" or "fucose-deficient" antibody variants include: US 2003/0157108; WO 2000/61739; WO 2001/29246; US 2003/0115614; US 2002/0164328; US 2004/0093621; US 2004/0132140; US 2004/0110704; US 2004/0110282; US 2004/0109865; WO 2003/085119; WO 2003/084570; WO 2005/035586; WO 2005/035778; WO2005/053742; WO2002/031140; okazaki et al, J.mol.biol.336:1239-1249 (2004); yamane-Ohnuki et al, biotech.Bioeng.87:614 (2004). Examples of cell lines capable of producing defucosylated antibodies include protein fucosylation deficient Lec13CHO cells (Ripka et al, arch. Biochem. Biophys.249:533-545 (1986); U.S. patent application No. US 2003/0157108 A1; and WO 2004/056312 A1, adams et al, particularly example 11), and knock-out cell lines, such as the alpha-1, 6-fucosyltransferase gene, FUT8 knock-out CHO cells (see, e.g., yamane-Ohnuki et al, biotech. Bioeng.87:614 (2004); kanda, Y. Et al, biotechnol. Bioeng.94 (4): 680-688 (2006); and WO 2003/085107).

Antibodies are also provided with bisected oligosaccharides, for example, where the biantennary oligosaccharides attached to the Fc region of the antibody are bisected by GlcNAc. Such antibody variants may have reduced fucosylation and/or improved ADCC function. Examples of such antibody variants are described, for example, in WO 2003/011878; U.S. Pat. nos. 6,602,684; and US 2005/0123546. Antibody variants having at least one galactose residue in an oligosaccharide linked to an Fc region are also provided. Such antibody variants may have improved CDC function. Such antibody variants are described, for example, in WO 1997/30087, WO 1998/58964 and WO 1999/22764.

Fc region variants

In certain examples, one or more amino acid modifications can be introduced into the Fc region of an antibody of the invention, thereby generating an Fc region variant. The Fc region variant may comprise a human Fc region sequence (e.g., a human IgG1, igG2, igG3, or IgG4 Fc region) comprising an amino acid modification (e.g., substitution) at one or more amino acid positions.

In certain instances, the disclosure contemplates antibody variants with some, but not all, effector functions, which make them desirable candidates for use where the half-life of the antibody in vivo is important and certain effector functions (such as complement and ADCC) are unnecessary or deleterious. In vitro and/or in vivo cytotoxicity assays may be performed to confirm the reduction/depletion of CDC and/or ADCC activity. For example, fc receptor (FcR) binding assays may be performed to ensure that the antibody lacks fcyr binding (and therefore may lack ADCC activity), but retains FcRn binding ability. The major cells mediating ADCC, NK cells, express Fc γ RIII only, whereas monocytes express Fc γ RI, fc γ RII and Fc γ RIII. FcR expression on hematopoietic cells is summarized in Table 3 on page 464 of ravatch and Kinet, annu.Rev.Immunol.9:457-492 (1991). Non-limiting examples of in vitro assays for assessing ADCC activity of a molecule of interest are described in U.S. patent nos. 5,500,362 (see, e.g., hellstrom, i.e., proc.natl.acad.sci.usa 83; U.S. Pat. No. 5,821,337 (see Bruggemann, M. Et al, J.Exp. Med.166:1351-1361 (1987)). Alternatively, non-radioactive assay methods may be employed (see, e.g., ACTI for flow cytometry) ^TM Non-radioactive cytotoxicity assay (CellTechnology, inc. Mountain View, CA); and CYTOTOX

Non-radioactive cytotoxicity assay (Promega, madison, WI)). Useful effector cells for such assays include Peripheral Blood Mononuclear Cells (PBMC) and autologous lymphocytesBut kill (NK) cells. Alternatively or additionally, the ADCC activity of the target molecule may be assessed in vivo, for example, in an animal model (such as disclosed in Clynes et al, proc.natl.acad.sci.usa 95-652-656 (1998)). A C1q binding assay may also be performed to confirm that the antibody is unable to bind C1q and therefore lacks CDC activity. See, e.g., the C1q and C3C binding ELISA in WO2006/029879 and WO 2005/100402. To assess complement activation, CDC assays can be performed (see, e.g., gazzano-Santoro et al, J.Immunol. Methods 202 (1996); cragg et al, blood.101:1045-1052 (2003); and Cragg et al, blood.103:2738-2743 (2004)). FcRn binding and in vivo clearance/half-life determinations can also be performed using methods known in the art (see, e.g., petkova et al, int' l. Immunol.18 (12): 1759-1769 (2006)).

Antibodies with reduced effector function include those with substitutions of one or more Fc region residues 238, 265, 269, 270, 297, 327 and 329 (U.S. Pat. nos. 6,737,056 and 8,219,149). Such Fc mutants include Fc mutants having substitutions at two or more of amino acid positions 265, 269, 270, 297 and 327, including so-called "DANA" Fc mutants having alanine substitutions for residues 265 and 297 (U.S. Pat. nos. 7,332,581 and 8,219,149).

Certain antibody variants with improved or reduced binding to FcR are described. ( See, e.g., U.S. Pat. nos. 6,737,056; WO 2004/056312; and Shields et al, J.biol.chem.9 (2): 6591-6604 (2001). )

In certain examples, an antibody variant comprises an Fc region with one or more amino acid substitutions that improve ADCC, e.g., substitutions at positions 298, 333, and/or 334 of the Fc region (EU numbering of residues).

In some cases, alterations are made in the Fc region, resulting in altered (i.e., improved or reduced) C1q binding and/or Complement Dependent Cytotoxicity (CDC), e.g., as described in U.S. Pat. Nos. 6,194,551, WO 99/51642 and Idusogene et al J.Immunol.164:4178-4184 (2000).

Antibodies with extended half-life and improved neonatal Fc receptor (FcRn) binding, responsible for the transfer of maternal IgG to the fetus (Guyer et al, J.Immunol.117:587 (1976); and Kim et al, J.Immunol.24:249 (1994)) are described in US2005/0014934A1 (Hinton et al). Those antibodies comprise an Fc region having one or more substitutions therein that improve binding of the Fc region to FcRn. Such Fc variants include those having substitutions at one or more of the following Fc region residues: 238. 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, 413, 424 or 434, for example, a substitution of residue 434 of the Fc region (U.S. Pat. No. 7,371,826).

For additional examples of Fc region variants, see also: duncan and Winter, nature322:738-40 (1988); U.S. Pat. nos. 5,648,260; U.S. Pat. nos. 5,624,821; and WO 94/29351.

Cysteine engineered antibody variants

In certain instances, it may be desirable to produce cysteine engineered antibodies, e.g., "thiomabs," in which one or more residues of the antibody are replaced with cysteine residues. In particular examples, the substituted residue is present at an accessible site of the antibody. As further described herein, the reactive thiol groups are positioned at accessible sites of the antibody by substituting those residues with cysteine, and can be used to conjugate the antibody to other moieties (such as a drug moiety or linker-drug moiety) to produce an immunoconjugate. In certain examples, cysteine can be substituted for any one or more of the following residues: v205 of the light chain (Kabat numbering); a118 of the heavy chain (EU numbering); and S400 of the heavy chain Fc region (EU numbering). Cysteine-engineered antibodies can be produced as described, for example, in U.S. Pat. No. 7,521,541.

Antibody derivatives

In certain examples, the antibodies provided herein can be further modified to contain additional non-proteinaceous moieties known in the art and readily available. Moieties suitable for derivatization of antibodies include, but are not limited to, water-soluble polymers. Non-limiting examples of water-soluble polymers include, but are not limited to, polyethylene glycol (PEG), copolymers of ethylene glycol/propylene glycol, carboxymethylcellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone, poly-1, 3-dioxolane, poly-1, 3, 6-trioxane, ethylene/maleic anhydride copolymers, polyamino acids (homopolymers or random copolymers) and dextran or poly (n-vinylpyrrolidone) polyethylene glycol, propylene glycol homopolymers, polypropylene oxide/ethylene oxide copolymers, polyoxyethylated polyols (e.g., glycerol), polyvinyl alcohol, and mixtures thereof. Polyethylene glycol propionaldehyde may have advantages in manufacturing due to its stability in water. The polymer may have any molecular weight and may or may not have branches. The number of polymers attached to the antibody can vary, and if more than one polymer is attached, they can be the same or different molecules. In general, the number and/or type of polymers used for derivatization can be determined based on considerations including, but not limited to, the particular property or function of the antibody to be improved, whether the antibody derivative will be used in therapy under defined conditions, and the like.

In another example, a conjugate of an antibody and a non-proteinaceous moiety that can be selectively heated by exposure to radiation is provided. In one example, the non-proteinaceous moiety is carbon nanotubes (Kam et al, proc.natl.acad.sci.usa 102. The radiation can be of any wavelength, and includes, but is not limited to, wavelengths that are not harmful to normal cells, but that heat the non-proteinaceous part to a temperature at which cells proximal to the antibody-non-proteinaceous part are killed.

Immunoconjugates

The invention also provides immunoconjugates comprising an antibody provided herein (e.g., an anti-PD-L1 antibody or an anti-PD-1 antibody) conjugated to one or more cytotoxic agents, such as a chemotherapeutic agent or drug, a growth inhibitory agent, a toxin (e.g., a protein toxin, an enzymatic activity toxin, or a fragment thereof, of bacterial, fungal, plant or animal origin), or a radioisotope.

In one instance, the immunoconjugate is an antibody-drug conjugate (ADC) in which the antibody is conjugated to one or more drugs, including but not limited to maytansinoids (see U.S. Pat. nos. 5,208,020, 5,416,064 and european patent EP 0 425 235 B1); auristatins (auristatins), such as monomethyl auristatin drug moieties DE and DF (MMAE and MMAF) (see U.S. Pat. nos. 5,635,483 and 5,780,588 and 7,498,298); dolastatin; calicheamicin or derivatives thereof (see U.S. Pat. Nos. 5,712,374, 5,714,586, 5,739,116, 5,767,285, 5,770,701, 5,770,710, 5,773,001, and 5,877,296 Hinman et al, cancer Res.53:3336-3342 (1993); and Lode et al, cancer Res.58:2925-2928 (1998)); anthracyclines, such as daunorubicin or doxorubicin (see Kratz et al, current Med. Chem.13:477-523 (2006); jeffrey et al, bioorganic & Med. Chem. Letters 16; methotrexate; vinblastine; taxanes such as docetaxel, paclitaxel, larotaxel, tesetaxel, and otaxel; trichothecene and CC1065.

In another instance, the immunoconjugate comprises an antibody described herein conjugated to an enzymatically active toxin or fragment thereof, including but not limited to diphtheria a chain, a non-binding active fragment of diphtheria toxin, exotoxin a chain (from pseudomonas aeruginosa), ricin a chain, abrin a chain, modeccin a chain, alpha-hypoxanthine, erythrin, dianthin protein, phytolacca americana protein (PAPI, PAPII, and PAP-S), momordica charantia inhibitor, curcumin, croton toxin, saporin inhibitor, gelatin, clindamycin (mitogellin), restrictocin, phenomycin, enomycin, and trichothecene.

In another example, an immunoconjugate comprises an antibody described herein conjugated to a radioactive atom to form a radioconjugate. A variety of radioisotopes are available for the production of radioconjugates. Examples include At ²¹¹ 、I ¹³¹ 、I ¹²⁵ 、Y ⁹⁰ 、Re ¹⁸⁶ 、Re ¹⁸⁸ 、Sm ¹⁵³ 、Bi ²¹² 、P ³² 、Pb ²¹² And radioactive isotopes of Lu. When the radioconjugate is used for detection, it may contain a radioactive atom for scintigraphic studies, for example tc99m or I123, or a spin label for Nuclear Magnetic Resonance (NMR) imaging (also known as magnetic resonance imaging, mri), such as iodine-123, iodine-131, indium-111, fluorine-19, carbon-13, nitrogen-15, oxygen-17, gadolinium, manganese or iron. Conjugates of the antibody and cytotoxic agent may be prepared using a variety of bifunctional protein coupling agents, such as N-succinimidyl-3- (2-pyridyldithio) propionate (SPDP), succinimidyl-4- (N-maleimidomethyl) cyclohexane-1-carboxylate succinimidyl ester (SMCC), iminothiolane (IT), bifunctional derivatives of imidoesters (such as dimethyl adipate hydrochloride), active esters (such as disuccinimidyl suberate), aldehydes (such as glutaraldehyde), bis-azido compounds (such as bis (p-azidobenzoyl) hexanediamine), bis-diazonium derivatives (such as bis- (p-diazoniumbenzoyl) -ethylenediamine), diisocyanates (such as toluene 2, 6-diisocyanate), and bis-active fluorine compounds (such as 1, 5-difluoro-2, 4-dinitrobenzene). For example, a ricin immunotoxin may be prepared as described in Vitetta et al, science 238 (1987). Carbon-14 labeled 1-isothiocyanatobenzyl-3-methyldiethylene triaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent for conjugating radionucleotides to antibodies. See WO94/11026. The linker may be a "cleavable linker" that promotes release of the cytotoxic drug in the cell. For example, acid-labile linkers, peptidase-sensitive linkers, photolabile linkers, dimethyl linkers, or disulfide-containing linkers can be used (Chari et al, cancer Res.52:127-131 (1992); U.S. Pat. No. 5,208,020).

Immunoconjugates or ADCs herein expressly contemplate, but are not limited to, such conjugates prepared with a cross-linking agent, including, but not limited to, commercially available (e.g., from Pierce Biotechnology, inc., rockford, il., u.s.a.) BMPS, EMCS, GMBS, HBVS, LC-SMCC, MBS, MPBH, SBAP, SIA, SIAB, SMCC, SMPB, SMPH, sulfo-EMCS, sulfo-GMBS, sulfo-KMUS, sulfo-MBS, sulfo-SIAB, sulfo-SMCC, sulfo-SMPB, and SVSB (succinimidyl- (4-vinylsulfone) benzoate).

Pharmaceutical compositions and formulations

The Pharmaceutical compositions and formulations described herein can be prepared in lyophilized formulations or in aqueous solution by mixing one or more active ingredients (e.g., anti-PD-L1 antibodies, such as atlizumab) of the desired purity with one or more optional Pharmaceutical carriers (Remington's Pharmaceutical Sciences 16 th edition, osol, a. Eds. (1980)). Pharmaceutically acceptable carriers are generally non-toxic to subjects at the dosages and concentrations used, and include, but are not limited to: buffers such as phosphates, citrates and other organic acids; antioxidants, including ascorbic acid and methionine; preservatives (such as octadecyl dimethyl benzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol, butanol or benzyl alcohol; alkyl parabens, such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents, such as EDTA; sugars such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions, such as sodium; metal complexes (e.g., zinc protein complexes); and/or a non-ionic surfactant, such as polyethylene glycol (PEG). Exemplary pharmaceutical carriers herein further include interstitial drug dispersants, such as soluble neutral active hyaluronidase glycoprotein (sHASEGP), e.g., human soluble PH-20 hyaluronidase glycoprotein, such as rHuPH20 ((r))

Baxter International, inc.). Certain exemplary sHASEGP and methods of use, including rHuPH20, are described in U.S. patent publication Nos. 2005/0260186 and 2006/0104968. In one aspect, the sHASEGP is combined with one or more additional glycosaminoglycanases (such as chondroitinase). It should be understood thatAny of the pharmaceutical compositions or formulations described above can include an immunoconjugate described herein instead of or in addition to a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

Exemplary lyophilized antibody formulations are described in U.S. Pat. No. 6,267,958. Aqueous antibody formulations include those described in U.S. Pat. No. 6,171,586 and WO 2006/044908, the latter formulations comprising histidine-acetate buffer.

The compositions and formulations herein may also contain more than one active ingredient necessary for the particular indication being treated, preferably active ingredients having complementary activities that do not adversely affect each other. For example, it may be desirable to further provide additional therapeutic agents (e.g., chemotherapeutic agents, cytotoxic agents, growth inhibitory agents, and/or anti-hormonal agents, such as those described above). Such active ingredients are suitably present in combination in an amount effective for the intended purpose.

The active ingredient may be embedded in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization (for example, hydroxymethylcellulose or gelatin-microcapsules and poly (methylmethacylate) microcapsules, respectively); embedded in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules); or embedded in the crude emulsion. Such techniques are disclosed in Remington's Pharmaceutical Sciences 16 th edition, osol, A. Eds (1980).

Sustained release preparations can be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the antibody, which matrices are in the form of shaped articles, e.g., films, or microcapsules. Formulations for in vivo administration are generally sterile. Sterility can be readily achieved by filtration, for example, through sterile filtration membranes.

Article and kit

In another aspect of the present disclosure, an article of manufacture or a kit comprising materials for treating, preventing, and/or diagnosing a subject is provided.

In some cases, such articles of manufacture or kits can be used to identify an individual having cancer (e.g., lung cancer (e.g., NSCLC), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), or breast cancer (e.g., TNBC)) who may benefit from a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attrituzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)). Such articles of manufacture or kits can include (a) reagents for determining the level of immune score expression of one or more of the genes in a sample from an individual, and (b) instructions for using these reagents to identify an individual having cancer (e.g., lung cancer (e.g., NSCLC), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), or breast cancer (e.g., TNBC)) who may benefit from treatment with a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., attritumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

For example, in some cases, an article of manufacture or kit can include (a) reagents for determining an immune score expression level of one or more genes listed in any one of tables 1 to 17 in a sample from an individual, and (b) instructions for using these reagents to identify individuals with cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) who may benefit from treatment with a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)).

In another example, provided herein is a kit comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attritumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) and instructions for administering the PD-L1 axis binding antagonist to an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) who has been identified as an individual who may benefit from treatment with the PD-L1 binding antagonist according to any of the methods disclosed herein.

In another example, provided herein is a kit comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, e.g., altlizumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)) and instructions to administer the PD-L1 axis binding antagonist to an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) for whom a treatment comprising the PD-L1 binding antagonist has been selected according to any one of the methods disclosed herein.

In another example, provided herein is a kit for identifying an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) who may benefit from treatment comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attrituzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), the kit comprising means for determining in a sample from the individual the expression levels of one or more of the genes CD79A, CD19, BANK1, jcain, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11) where the immune expression level of the one or more of the gene(s) would benefit from a high level of the identification of the PD-L1 axis binding antagonist in the individual.

In another example, provided herein is a kit for identifying an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) who may benefit from treatment comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attrituzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), the kit comprising means for determining the expression level of one or more of genes CD79A, CD19, BANK1, jcain, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein the individual is identified as a PD-L1 axis binding antagonist that may comprise a PD-L1 axis binding antagonist if the immune score expression level of the one or more genes is higher than a reference immune score expression level of the one or more genes, wherein the individual does not benefit from treatment comprising prolonged PD-L1 axis binding to the individual.

In some cases, the kit comprises reagents for determining the expression level of two or more of genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual. In some cases, the kit includes reagents for determining the expression levels of three or more of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual. In some cases, the kit includes reagents for determining the expression levels of four or more of genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual. In some cases, the kit includes reagents for determining the expression level of five or more of genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from an individual. In some cases, the kit comprises reagents for determining the expression level of six or more of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual. In some cases, the kit comprises reagents for determining the expression level of seven or more of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual. In some cases, the kit comprises reagents for determining the expression level of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the subject.

The kit may include reagents for determining the expression level of any combination of B cell signature genes. For example, the combination may include two genes selected from CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1, e.g., any one of the combinations listed in table 3. In another example, the combination can include three genes selected from CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1, e.g., any one of the combinations listed in table 4. In another example, the combination can include four genes selected from CD79A, CD19, BANK1, jchan, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1, e.g., any one of the combinations listed in table 5. In another example, the combination can include five genes selected from CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1, e.g., any one of the combinations listed in table 6. In another example, the combination can include six genes selected from CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1, e.g., any one of the combinations listed in table 7. In another example, the combination may include seven genes selected from CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1, e.g., any one of the combinations listed in table 8.

In another example, provided herein is a kit for identifying an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) who is likely to benefit from treatment comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as astuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), the kit comprising means for determining in a sample from the individual one or more of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 13, 14, or more of the genes are expressed at a high immune score for the individual, wherein the individual is likely to benefit from expression of the reference level of the one or more of the individual.

In another example, provided herein is a kit for identifying an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) who may benefit from treatment comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attrituzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), the kit comprising reagents for determining the expression levels of one or more of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from the individual, wherein a reference level of immune expression of the one or more genes at which is higher than the reference level of immune expression of the one or more genes would benefit from treatment comprising an extended PD-axis binding to the individual, wherein the subject would benefit from treatment using the PD-L1 axis binding antagonist.

In some cases, the kit includes reagents for determining the expression level of two or more of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from an individual. In some cases, the kit comprises reagents for determining the expression level of three or more of the genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from an individual. In some cases, the kit includes reagents for determining the expression level of four or more of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from an individual. In some cases, the kit comprises reagents for determining the expression level of five or more of the genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from an individual. In some cases, the kit comprises reagents for determining the expression level of six or more of the genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from an individual. In some cases, the kit comprises reagents for determining the expression level of seven or more of the genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from an individual. In some cases, the kit includes reagents for determining the expression level of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from an individual.

The kit may include reagents for determining the expression level of any combination of plasma B cell signature genes. For example, the combination may include two genes selected from MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5, e.g., any one of the combinations listed in table 10. In another example, the combination may include three genes selected from MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5, e.g., any one of the combinations listed in Table 11. In another example, the combination may include four genes selected from MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5, e.g., any one of the combinations listed in table 12. In another example, the combination may include five genes selected from MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5, e.g., any one of the combinations listed in table 13. In another example, the combination may include six genes selected from MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5, e.g., any one of the combinations listed in table 14. In another example, the combination may include seven genes selected from MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5, e.g., any one of the combinations listed in table 15.

In another example, provided herein is a kit for identifying an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) who may benefit from treatment with a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attritumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), the kit comprising reagents for determining the presence of TLS in a tumor sample from the individual, wherein the presence of TLS in the tumor sample identifies the individual as an individual who may benefit from treatment with a PD-L1 axis binding antagonist.

In another example, provided herein is a kit for identifying an individual having a cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., NSCLC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) who may benefit from treatment comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as alemtuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), the kit comprising a means for determining the expression level of one or more of genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12) in a sample from the individual, wherein the immune expression level of the one or more genes is scored as high as a reference level of the individual's potential binding to benefit from the reference PD 1.

In some cases, a kit comprises reagents for determining the expression level of two or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from an individual. In some cases, a kit comprises reagents for determining the expression levels of three or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from an individual. In some cases, a kit comprises reagents for determining the expression levels of four or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from an individual. In some cases, a kit comprises reagents for determining the expression level of five or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from an individual. In some cases, a kit comprises reagents for determining the expression level of six or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from an individual. In some cases, a kit comprises reagents for determining the expression level of seven or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from an individual. In some cases, a kit comprises reagents for determining the expression level of eight or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from an individual. In some cases, a kit comprises reagents for determining the expression level of nine or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from an individual. In some cases, a kit comprises reagents for determining the expression level of ten or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from an individual. In some cases, a kit comprises reagents for determining the expression level of eleven or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from an individual. In some cases, a kit comprises reagents for determining the expression level of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from an individual.

In another example, provided herein is a kit for identifying an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) who may benefit from treatment comprising a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as atuzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), the kit comprising determining the number of B cells in a tumor sample from the individual, wherein the individual is identified as an individual who may benefit from treatment comprising a PD-L1 axis binding antagonist if the number of B cells in the tumor sample is greater than a reference number of B cells.

In another example, provided herein is a kit for identifying an individual having cancer (e.g., lung cancer (e.g., NSCLC)), bladder cancer (e.g., UC), kidney cancer (e.g., RCC), and breast cancer (e.g., TNBC)) who may benefit from treatment with a PD-L1 axis binding antagonist (e.g., a PD-L1 binding antagonist (e.g., an anti-PD-L1 antibody, such as attrituzumab) or a PD-1 binding antagonist (e.g., an anti-PD-1 antibody)), the kit comprising reagents for determining whether the individual has clonally expanded B cells in a tumor sample from the individual, wherein the clonally expanded B cells in the sample identify the individual as an individual who may benefit from treatment with a PD-L1 axis binding antagonist.

Either of the articles or kits may further comprise carrier means compartmentalized to receive within a well-defined space one or more container means (such as vials, tubes, etc.), wherein each of the container means comprises one of the individual elements to be used in the method. In the case of articles or kits for detecting target nucleic acids using nucleic acid hybridization, the kit may also have a container containing nucleotides for amplifying the target nucleic acid sequence and/or a container containing a reporter molecule such as an enzymatic, fluorescent, or radioisotope label.

In some cases, the article of manufacture or kit comprises the above-described container and one or more other containers comprising materials desirable from a commercial and user standpoint, including buffers, diluents, filters, needles, syringes, and package inserts with instructions for use. A label may be present on the container to indicate that the composition is for a particular application, and may also indicate guidelines for in vivo or in vitro use, such as those described above. For example, the article of manufacture or kit may further comprise a container comprising a pharmaceutically acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate buffered saline, ringer's solution, and dextrose solution.

The articles of manufacture or kits described herein can have many embodiments. In one instance, an article of manufacture or a kit comprises a container, a label on the container, and a composition contained within the container, wherein the composition comprises one or more polynucleotides that hybridize under stringent conditions to a complement of a gene listed herein (e.g., one or more genes listed in any one of tables 1-17), and the label on the container indicates that the composition can be used to assess a sample for the presence of a gene listed herein (e.g., one or more genes listed in any one of tables 1-17), and wherein the kit comprises instructions for using the one or more polynucleotides for assessing the presence of gene RNA or DNA in a particular sample type.

For oligonucleotide-based articles or kits, the article or kit can include, for example: (1) An oligonucleotide, e.g., a detectably labeled oligonucleotide, that hybridizes to a nucleic acid sequence encoding a protein, or (2) a pair of primers useful for amplifying a nucleic acid molecule. The article of manufacture or kit may also include, for example, a buffer, a preservative, or a protein stabilizing agent. The article of manufacture or kit may further comprise components necessary for the detection of a detectable label (e.g., an enzyme or substrate). The article of manufacture or kit can further comprise a control sample or a series of control samples that can be assayed and compared to the test sample. Each component of the article of manufacture or kit can be enclosed within a separate container, and all of the various containers and instructions for interpreting the results of the assays performed using the kit can be in separate packages.

In one instance, an article of manufacture or kit comprises a container, a label on the container, and a composition contained within the container, wherein the composition comprises one or more antibodies that specifically bind under stringent conditions to any of the biomarkers disclosed herein (e.g., one or more genes, TLS, or B cells (including clonally expanded B cells) listed in any one of tables 1 to 17), and the label on the container indicates that the composition can be used to assess the presence of the biomarkers listed herein in a sample, and wherein the kit comprises instructions for using the one or more antibodies for assessing the presence of a biomarker in a particular sample type.

VII. examples

The following are examples of the process of the present invention. It is to be understood that various other embodiments may be practiced given the general description provided above.

Example 1 tumor infiltrating B cells (including plasma B cells) and tertiary lymphoid structures are correlated with tumor response to PD-L1 blockade in NSCLC

Unsupervised differential analysis of gene expression was performed in pre-treatment biopsies of the phase 2 POPLAR trial (NCT 01903993). The study confirmed the results of phase 3 OAK (NCT 02008227) and BIRCH (NCT 02031458) clinical trials. This analysis, described in more detail in this example, reveals a strong correlation between the presence of B cells (particularly plasma B cells) and Tertiary Lymphoid Structures (TLS) in a patient's tumor and the continued manifestation of the sustained survival benefits associated with atezumab.

Unsupervised differential gene expression analysis showed that the B cell genes were associated with Overall Survival (OS) following alemtuzumab treatment

Whole genome RNA sequence data from archived tumor samples from POPLAR trial patients were analyzed. Gene expression data for patients with <6 months of survival (n = 24) receiving treatment with atelizumab and patients with > 12 months of survival (n = 43) were compared (Lee, J. Et al, reinvigorating activated T Cells by Block of the PD-1 pathway for Immunopathol. Dis. Therap.6,7-17 (2015)). Unsupervised differential gene expression analysis showed that, after correction of multiple hypothesis tests, multiple B cell genes were significantly associated with OS in addition to the expected T cell effector genes (fig. 1A and 7).

To assess whether these B cell gene signatures (CD 79A, SLAMF7, BTK, TNFRSF17, IGJ or jcain, IGLL5, RBPJ and MZB 1) were associated with OS in both treatment groups, the mean z scores for these genes were associated with benefit in the alemtuzumab group (greater than median, HR =0.39, ci =0.22-0.68, p < <0.001) and less relevant with chemotherapy or docetaxel treatment (HR =0.74, ci =0.52-1.07, p =0.1 relative to docetaxel) (fig. 1B). Among all these genes, the B cell receptor gene (CD 79A) was one of the most significantly differentially expressed genes, and high expression of CD79A was associated with OS benefit in the alemtuzumab-treated group (greater than median, HR =0.36, ci =0.2-0.64, p < -0.001) (fig. 1C).

B cell transcriptional signature was associated with prolonged OS in patients receiving treatment with atelizumab

Attempts were also made to identify Differentially Expressed Genes (DEG) between tumors in astuzumab responders and non-responders in OAK, comparing patients with long survival (> 12 months, n = 313) versus patients with short survival (< 6 months, n = 205) within each treatment group. 817 genes are differentially expressed between patients with long OS or short OS who received treatment with astuzumab (P <0.01 after FDR correction, absolute logFC ≧ 0.5). Genes with high degrees of overexpression were enriched in genes associated with B and plasma cell biology, including CD19, CD79A, BANK1, JCHAIN, MZB1, and TNFRSF17 (BCMA) (fig. 15A). Genes associated with cytotoxic T cell and IFN signaling (CD 3E, D, G, CD8A, GZMA-B, IFNG, CXCL 9-10) were also detected, but not significantly. None of these genes were significant in the docetaxel group (fig. 15B), indicating that these B/plasma cell genes have particular predictive value for the response to alemtuzumab. These observations were validated in phase II randomized clinical trial POPLAR (fig. 21A and 21B).

The results of the linear modeling were then verified by deriving a Kaplan-Meier curve in OAK, classifying gene expression by the tertile and comparing combined high (T3) and low/moderate (T1-T2) expression. The B cell markers CD79A (HR alemtuzumab: 0.54[ 2] 0.41-0.72]; HR chemotherapy: 0.88[0.67-1.13 ]) and CD19 (HR alemtuzumab: 0.65[0.50-0.86]; HR chemotherapy: 0.94[0.72-1.2 ]) provide an increased OS benefit as compared to IFNG (HR alemtuzumab: 0.73[0.55-0.96]; HR chemotherapy: 0.95[0.73-1.23 ]) or IFN-inducible chemokine CXCL10 (HR alemtuzumab: 0.85[0.65-1.11]; HR chemotherapy: 1.11[0.85-1.44 ]) (FIG. 15C-15F). Also, similar results were observed in POPLAR (fig. 21C to 21F).

To confirm these transcriptional findings at the protein level, baseline tumor samples from two attritumab responders and two attritumab non-responders were evaluated by Immunofluorescence (IF) staining for CD8, CD79, and Ki67 using multiple IFs. Abundant CD79+ B cells were detected in tumors from patients with objective remission compared to patients without objective remission (fig. 15G). In addition, multiple Immunohistochemistry (IHC) detected regions in which CD8+ T cells were immediately adjacent (9.9 ± 1.69 μm apart) to CD79+ B cells, suggesting that there was B cell-T cell interaction.

Identification of B cell subpopulations by Single cell RNA-seq

To further characterize the B-cell compartments involved in response to atuzumab, a large scRNA-seq dataset of 208,506 cells collected from lung and peripheral tissues of 44 NSCLC patients was analyzed with a focus on the B/plasma cell population. Cells were aggregated from tumor and draining lymph node compartments, dimensionality reduction was performed using Unified Manifold Approximation and Projection (UMAP) and graph-based clustering, identifying three major CD79A + B cell subsets in tumors, including follicular B cells, germinal Center (GC) B cells, and plasma cells (fig. 16A). Although the number of B/plasma cells was found to be increased in tumor tissue compared to Normal Adjacent Tissue (NAT), the composition of GC, follicular B and plasma cells within these B cell populations was similar between tumors and NAT from the same patient (fig. 16B). Transcription characteristics unique to each subset are identified. Follicular B cells are rich in CD83, CD69, SELL and BANK1 (Aiba et al, 2006). Plasma cells are rich in several immunoglobulin transcripts (ighg2, 137ighgp, IGHA 2), MZB1, DERL3 and XBP1. Germinal center B cells are rich in HMGA1, HMGB2 and RGS13. These characteristics were refined by exploring the expression of these transcripts in various immune cells (bone marrow, NK and T cells), stroma (fibroblasts, endothelial cells) and epithelial/tumor cell populations from the same patient (fig. 22A), retaining only B-cell compartment specific genes. Further focus was on only markers with high specificity for a single B cell population (fig. 16C and 22A).

The transcription method was supplemented with mass cytometry using a CyTOF combination containing 38 markers for single cell suspensions from six purchased NSCLC tumors. The analysis confirmed the presence of three intratumoral B cell populations based on CD19, HLA-DR, CD38 and Ki67 staining (plasma cells: CD38+ +, HLA-DR-, ki67-; GC B cells: CD38+, HLA-DR +, ki67+; follicular B cells: CD38-, HLA-DR +, ki 67-) (FIG. 16D). These specific markers were also verified to have similar expression patterns in the scRNA-seq dataset (FIG. 22B). In summary, single cell analysis identified three major intratumoral B cell subsets with highly specific markers.

High plasma cell characteristics predict OS benefit from alemtuzumab

Then, it was investigated whether these B-cell characteristics identified according to the scRNA-seq analysis could predict the survival of patients after receiving treatment with attrituximab or docetaxel. First, by ensuring gene co-expression through hierarchical clustering (fig. 17A) and intergenic correlation (fig. 23), it was verified that scRNA-seq derived features can be applied to bulk RNA-seq data. These analyses showed that there was a high correlation between germinal center B cells and follicular B cells in a large number of tumors (fig. 17b, r = 0.74), whereas tumors with high plasma cells were more pronounced. Plasma, GC B and follicular B cell characteristics are summarized in table 18.

Table 18: follicular B cell, GC B cell, and plasma B cell gene signatures

Follicular B cells	GC B cells	Plasma cells
			BANK1	GCSAM	MZB1
LINC00926	LRMP	DERL3
			FCER2	AICDA	JSRP1
GAPT	AC023590.1	TNFRSF17
			HVCN1	SUSD3	SLAMF7
		IGHG2
					IGHGP
		IGLV3-1
					IGLV6-57
		IGHA2
					IGKV4-1
		IGKV1-12
					IGLC7
		IGLL5

These features were then bisected using a three-place score split and comparing groups expressing high (T3) with low/medium (T1/T2). Kaplan-Meier analysis showed that although a tendency of OS benefit was observed with atuzumab in three characteristics, only plasma cell characteristics were significant (atuzumab high vs. low HR =0.63, 0.48-0.83]; chemotherapy high vs. low HR =0.92, 0.71-1.2; atuzumab high vs. chemotherapy high HR =0.64, 0.47-0.88) (FIGS. 18A-18D). In addition, tumors with high plasma cells showed significantly more patients experiencing complete remission, partial remission, or optimal overall remission with sustained stable disease (SD ≧ 6 months) (FIG. 24A). In POPLAR, there was also similar significant stratification of plasma cell characteristics for overall survival (fig. 24B-24D). The predictive value of plasma cell characteristics was confirmed in the model that tested the interaction of each characteristic with the treatment group (fig. 18E). Importantly, GC B cells, follicular B cells and our 8 gene T effector signature tGE8 (consisting of IFNG, CXCL9, CD8A, GZMA, GZMB, CXCL10, PRF1, TBX 21) were not significant in these models (fig. 18E). The predicted values of plasma cell characteristics were also present in a multivariate model including all four characteristics (atlizumab HR =0.67, [0.50-0.89]; chemotherapy HR =0.94, [0.70-1.26 ]), confirming that the effects observed with plasma cell characteristics are specific and independent of the presence of CD 8T cells (FIG. 18F). Increased B cells have been described as having prognostic significance for TCGA, among other indications; however, our data indicate that NSCLC tumors rich in plasma cells have strong predictive signatures, particularly for immune checkpoint inhibition. Indeed, plasma cell characteristics split by the three-score split method in TCGA NSCLC tumors had no effect on stratification of overall survival (fig. 24E). Taken together, these data indicate that plasma cell-rich tumors are predictive of overall survival benefit in patients receiving specific immune checkpoint inhibitor therapy.

Enriched plasma cell characteristics in tumors with tertiary lymphoid structures and/or lymphoid aggregates

In POPLAR, tumor hematoxylin and eosin (H & E) slides were evaluated for the presence of only TLS or lymphoid aggregates (LA, no germinal centers observed) (fig. 19A). Of the 254 patient samples analyzed, 9% had TLS-like structures with germinal centers, 21% had lymph aggregates (no germinal centers observed), and the remaining 70% had no detectable TLS-like or lymph aggregates. Lymph node metastasis samples (4% of all samples) were excluded from the analysis (fig. 19B). Patients with tumors containing TLS and/or LA showed a significant increase of 191OS (HR =0.60 2 2.38-0.94) when receiving treatment with atelizumab, but not when receiving treatment with docetaxel (HR =0.93 2-0.1.41) (fig. 19C).

A linear model was then applied to identify DEG between TLS/LA + and TLS/LA-tumors. 928 genes were differentially expressed between the two groups (P <0.01 after FDR correction, absolute logFC > 0.5). TLS/LA + tumors were highly enriched with genes from three intratumoral B cell subsets, but especially plasma cell genes (including MZB1, TNFRSF17 (BCMA) and immunoglobulins (fig. 20A and 20B.) quantitative analysis of the three B cell characteristics confirmed an increase in plasma cells (P =4.1 e-12) in TLS/LA + tumors as well as GC B cells (P =2.6 e-07) and follicular B cell characteristics (P =5.3 e-05) (fig. 20C).

In summary, our study integrated batch transcriptome and single cell RNA and protein measurements from two large randomized clinical trials of attrituzumab and docetaxel, demonstrating that baseline plasma cell enrichment in NSCLC tumors is an effective predictor of OS benefit following immune checkpoint blockade.

The correlation of clinical benefit of B cells after treatment with astuzumab was confirmed in an independent clinical trial dataset

To confirm the above results, two independent clinical data sets, i.e. BIRCH (n = 591) and OAK (n = 727), were analyzed. Interestingly, in the randomized phase 3 OAK study, high expression of B cell characteristics was associated with OS benefit in the chemotherapy and alemtuzumab treated groups, with greater benefit observed in the alemtuzumab treated group (fig. 2A). These data indicate that high expression of B cell features may have both prognostic and predictive components for OS (greater than median, HR =0.84, ci =0.71-1, p =0.056 relative to docetaxel, and HR =0.54, ci =0.42-0.7, p <0.001 relative to the less-than-median B cell gene set within the attritumab group). Similarly, in phase 3 BIRCH studies with single-therapy single-panel PD-L1 selection (immune or tumor cell positive rate greater than 5%), high expression of B cell signatures was found to be associated with OS benefit in patients receiving treatment with atelizumab (HR =0.78, ci =0.62-0.99, p = 0.044) (fig. 2B).

The best-confirmed overall response (BCOR) and progression-free survival (PFS) in the OAK study were also evaluated. Responders to alemtuzumab showed significantly higher B-cell signature gene expression than non-responders, and this difference was not observed in patients receiving docetaxel treatment (fig. 2C). Similar to the association with OS, high expression of B cell gene signatures was associated with greater PFS benefit in both treatment groups, with greater benefit observed in the alemtuzumab group (greater than median, HR =0.54, ci =0.42-0.7, p < -0.001) or relative to docetaxel (HR =0.84, ci =0.71-1, p = 0.056) (fig. 2D). High B cell characteristics were associated with greater benefit of lung tumors (except lymph node metastases), regardless of histology or sample type (biopsy or resection) (fig. 14A to 14C).

Confirming the presence of B cells in a NSCLC sample from a patient

To confirm whether these gene expression results in the presence of CD79+ B cells in NSCLC tumors, CD8, CD79 and Ki67 in baseline tumor samples from 20 patients were evaluated by IF staining using multiple Immunofluorescence (IF). These patient samples had high or low CD79A gene expression (fig. 3A). Abundant CD79+ B cells were detected in NSCLC samples with high CD79A RNA. Furthermore, multiple Immunohistochemistry (IHC) detected regions in which CD8+ T cells were in close proximity (9.9 ± 1.69 μm apart) to CD79+ B cells, suggesting that there was B cell-T cell interaction.

The association of the B cell gene CD79A with OS is independent of the T cell gene

The presence of T cells in the tumor microenvironment can up-regulate cytokines and chemokines that non-specifically recruit other immune cells. To assess whether B cells present in a tumor are likely bystander cells recruited into the tumor due to infiltrating T cells, and given that immune cell genes are generally highly correlated (in our data set, r for CD79A and CD8A, for example) ² =0.58, r for CD3D and CD79A ² =0.61, using Cox proportional airMultivariate regression analysis at risk to assess whether the association of CD79A (B cell receptor gene) is dependent on CD3D, CD3E, CD8A gene and T _eff The presence of genetic characteristics. This method has been used to assess tumor-infiltrating lymphocyte (TIL) anatomy in colorectal cancer (CRC) and novel immune characteristics in hepatocellular carcinoma (HCC) (Berntsson, J. Et al, the clinical impression of tumor-infiltrating lymphocytes in clinical cancer by cancer subsidiary sub site: A clinical study. Int. J. Cancer 141,1654-1666 (2017); tian, M.X. Et al, tissue-infiltrating lymphocytes signature prediction in Patients with early/interface Tissue specimen clinical specimen in cancer Carcinoma. BMC.20117, 106 (2019)). Using Cox proportional Risk multivariate regression analysis, in the analysis of CD3D, CD3E, CD8A genes or T _eff The association of CD79A with OS was still statistically significant after gene expression was controlled as a covariate (table 19). This indicates that expression of the CD79A gene at baseline has a significant impact on OS risk ratio in patients receiving alemtuzumab treatment, independent of T cell genes in the tumor microenvironment.

Table 19: statistical analysis associated with Cox ratio risk survival analysis comparing CD79a and CD3 genes from POPLAR

Correlation with OS compared to CD3D

Likelihood ratio test =23.9 (on 2 df), p =6.44e-06, n =92, event number =51

Correlation with OS compared to CD3E

Likelihood ratio test =23.9 (at 2 df), p =6.44e-06, n =92, number of events =51

Correlation with OS compared to CD8A

Likelihood ratio test =23.45 (at 2 df), p =8.106e-06, n =92, event number =51

Compared with T _eff Features, dependency on OS

Likelihood ratio test =23.95 (at 2 df), p =6.298e-06, n =93, number of events =52

High CD79A + tumors show the presence of Tertiary Lymphoid Structures (TLS)

To better understand the role of B cells in these tumors, tumor hematoxylin and eosin (H & E) slides were evaluated for the presence of TLS (fig. 3B). TLS is often observed in tissues affected by chronic inflammation caused by infection, autoimmunity, cancer and allograft rejection. These highly ordered structures, similar to the cellular composition of the lymphoid follicles, are thought to mimic the activity of germinal centers and contribute to the local control of the adaptive immune response. Of the 290 NSCLC patient samples analyzed, 31% had TLS with germinal centers, 30% had lymphatic aggregates (no germinal centers observed), and the remaining 39% had no detectable TLS or lymphatic aggregates. Lymph node metastasis samples (4% of all samples) were excluded from the analysis. In this cohort, TLS and lymphatic aggregates of squamous and non-squamous NSCLC types were similarly distributed and were mainly detected in the resection samples (fig. 9A and 9B). Importantly, tumors with high CD79A gene expression were highly correlated with the presence of TLS (fig. 9C).

Although approximately equal numbers of TLS were detected in tumors of both treatment groups, TLS-positive patients treated with atuzumab showed an association with OS benefit, which was not observed in TLS-positive patients treated with docetaxel in the POPLAR (fig. 3E) and OAK studies (fig. 9F).

To further confirm that these Structures identified by H & E staining are indeed TLS, a subset of the samples were subjected to IHC to detect specific vasculature or High Endothelial Venules (HEV) known to associate with TLS and mediate lymphocyte trafficking to secondary Lymphoid organs (Colbeck, E.J. et al, therapeutic lymphoma Structures in Cancer: drivers of anticancer, immunosuppresion, or Bystandander Sentists in DisaseFront.8, 1830 (2017); engelhard, V.H. et al, immune Cell induction and therapeutic lymphoma Structures as Determinants of anticancer Immunity.J.200, 432-442 (2018)). The MECA-79 monoclonal antibody recognizes peripheral lymph node addressen (PNAd) by targeting CD62L or L-selectin expressed exclusively by HEV. Tumor samples from TLS positive patients were positive for MECA-79, CD40 and CD8 by IHC staining (FIGS. 4A to 4D).

TLS gene signature associated with Attuzumab OS benefit

As described above, by H&E TLS analysis was performed on a limited number (n = 290) of samples. To extend this analysis to the larger gene expression dataset we have available, previously reported TLS gene signatures were used. Confirmation based on H&The TLS sample of E was associated with the published TLS gene expression profiles (FIG. 9E), and also with the B-cell gene profile and T _eff Gene signature correlated with expected results (FIGS. 9C and 9D) (Zhu, G. Et al, tumor-Associated temporal identification Structures: gene-Expression Profiling and Their bioengineering. Front. Immunological.8, 767 (2017); colbeck, E.J. et al, temporal localization Structures in Cancer: drivers of anti-Tumor Immunity, or Bystander Senttinels in DisaseFront.Immunol.8, 1830 (2017), bremenes, R.M. et al, the Role of Tumor-embedding pathologies in Development, progress, and progress of Non-Cell luminescence cancer.J. third organic. Oncol.11,789-800 (2016; german, C. Et al, presence of B Cell in Tertiary localization Structures with a property of a tissue in Cancer J.2014.2014.42, mass. C. Et al, presence of B Cell in Tertiary localization Structures in Cancer with a property of tissue J.Oncol.2014.2014.2014).

Patient tumors with high TLS gene signature were significantly correlated with OS benefit in the atlizumab treatment group in POPLAR (greater than median, HR =0.48, ci =0.27-0.86, p = 0.013) and in the oatlizumab treatment group in OAK (greater than median, HR =0.82, ci =0.64-1.06, p = 0.14) (fig. 5A and 5B). Similarly, using another TLS-associated gene signature identified by Cabrita et al, the hair-center initiation signature shows that treatment with atuzumab in POPLAR (greater than median, HR =0.44, ci =0.23-0.84, p = 0.013) and in OAK (greater than median, HR =0.66, ci =0.51-0.86, p = 0.01) showed significant survival benefits (fig. 5C and 5D) (Cabrita, r. Et al, terrial simple temporal structure improved survival methodology and survival efficiency in tumor 577,561-565 (2020)).

Plasma cell gene signature associated with astuzumab OS benefit

To further elucidate the B-cell subtypes present in tumors, several known B-cell characteristics were analyzed for their ability to predict survival (fig. 11A to 11C) (Newman, a.m. et al, robust expression of cell subsets from tissue expression profiles. Nat. Methods 12,453-457 (2015)). OS benefit treated with Attributumab shows one of the strongest associations with published Plasma Cell characteristics consisting of the double gene set TNFRSF17 (or BCMA) and the immunoglobulin junction region (IGJ or JCHAIN) common to all antibodies (Kroeger, D.R. et al, tumor-Infiltrating Plasma Cell array Associated with therapeutic approach Structures, cytolytic T-Cell Responses, and Superior Prognosis in Ovarian cancer. Clin. Cancer Res.22,3005-3015 (2016)). This high expression of the dual-gene plasma cell signature was closely correlated with OS benefit in OAKs with treatment with atuzumab (greater than median, HR =0.58, ci =0.44-0.74, p-straw 0.001) (fig. 6A).

Reduction of the Shannon Diversity Index (SDI) of the BCR (B cell receptor) lineage is associated with disease control

Given that plasma cell characteristics are highly correlated with clinical benefit, we wanted to investigate if there were any changes in BCR lineage or clonality following alemtuzumab treatment in patients exhibiting clinical benefit. 13 pairs of matched pre-and post-treatment biopsy samples from the alemtuzumab monotherapy treatment FIR (NCT 01846416) and BIRCH trials were analyzed as shown in fig. 6A to 6E. All post-treatment samples were taken at the time of disease progression. Median acquisition time was 184.5 days after initial treatment for patients with Partial Remission (PR) or patients with Stable Disease (SD); for patients with Progressive Disease (PD), median acquisition time was 51 days after initial treatment. The only PR patients analyzed here had a PFS of 12.9 months. 7 patients with SD and 5 patients with PD had median PFS of 5.52 months and 1.35 months, respectively. Due to the deletion, neither group reaches the median OS. Tumor RNA was isolated and BCR gene was sequenced. RNA-based clonal lineage Sequencing enables us to determine isotypes and exclude signal dilutions of non-expressing B cells (Schalper, K.A. et al, neoadjuvant nivolumab modifications the Tumor immunity in responsive leukemia in. Nat.Med.25,470-476 (2019); smith, C.C. et al, using RNA Sequencing of the Characterise the Tumor immunity. Methods mol. Biol.2055,245-272 (2020)). Most BCRs sequenced were of the IgG isotype, not IgA or IgM, indicating that tumor infiltrating B cells were predominantly IgG + B cells (fig. 12A to 12F).

Once the BCR cDNA library is generated and sequencing is complete, the BCR lineage will be evaluated using SDI (a measure of B cell clonality) (Chaudhary, N. And Wesemann, D.R. analytical immunology reproduction. Front. Immunol.9,462 (2018); greiff, V. Et al. A Bioformatic frame for immunological specificity profiling purposes detection of immunological status. Genome. Med.7,49 (2015); kaplinky, J. Et al. Butyl esters of immunological specificity samples on samples. Nat.7, 81 (Comm. 1181187)). SDI of BCR sequences of pre-and post-alemtuzumab treatment pairs (n = 11) of NSCLC patient tumor samples were compared to pre-treatment tumor sample pairs (n = 3), where both samples were taken before treatment began as experimental controls. Paired samples taken from 3 patients before treatment with atezumab showed no change in SDI (fig. 6B). Among the patients analyzed for the pre-and post-alemtuzumab samples, 7 patients with SD and 1 patient with PR showed a significant decrease in the shannon index of the tumor after treatment compared to the pre-treatment samples (p =0.0078, fig. 6C). In contrast, the post-treatment index was not significantly changed in 5 patients with PD (p = 0.8125) (fig. 6D). These observations indicate that BCR diversity is reduced in this limited subset of patients who show clinical benefit (PR and SD) for atelizumab. This indicates that clonal expansion or enrichment of specific Ig + B cells occurs in tumors treated with astuzumab. 5 patients receiving alemtuzumab treatment showed no evidence of clonal enrichment during treatment (fig. 6E).

Discussion of the preferred embodiments

Our data indicate that infiltration of B cells (specifically surface Ig + B cells and plasma cells) into the tumor microenvironment is a key factor in determining OS benefit in NSCLC patients receiving treatment with atelizumab. Our data demonstrate that the presence of B cells and TLS at baseline determines the atelizumab-mediated OS benefit, and these results were validated in three large independent clinical trial cohorts (one phase 2 study [ n =193] and two phase 3 studies [ n =591 and 727], respectively).

In our study, gene expression analysis showed a strong association between B-Cell markers (memory Cells and Plasma Cells) and the survival benefit of alemtuzumab treatment (fig. 1A to 1C, fig. 2A to 2D, and fig. 11A to 11C), and includes the Plasma Cell gene characteristics of BCMA and IGJ genes (Kroeger, d.r. et al, tumor-profiling Plasma Cells electrode Associated with clinical laboratory Structures, cytolytic T-Cell Responses, and Superior Prognosis in Ovarian cancer. Clin. Cancer res.22,3005-3015 (2016)). Data from BCR sequencing showed a decrease in intratumoral B cell diversity using SDI in patients showing clinical benefit to atelizumab and indicated the presence of B cell clonal expansion.

Analysis showed that CD79+ B cells were associated with CD8 when visualized by immunofluorescence imaging ⁺ The close proximity of T cells provides support for the role of these cells as APCs.

In this study, a strong correlation between CD79A gene expression and the presence of TLS was observed, as determined by H & E staining. TLS in NSCLC is reported to be predominantly present in early-stage Squamous NSCLC patients (> 95%), with 50% of patients having no Germinal center (Silina, K. et al, germinal center determination of the cosmetic Relevance of clinical laboratory Structure and article affected by clinical society in Lung Squamous Cell Carcinoma. Cancer Res.78,1308-1320 (2018)). Analysis of metastatic NSCLC showed that the prevalence of TLS was about 30%, and another 30% of patients had lymphatic aggregates without germinal centers. Our analysis also showed the presence of TLS histologically and gene expression profiles showed significant correlation of OS after atuzumab treatment, suggesting that they may play an important role in PD1/PD-L1 blocking the sustained immune response of tumors.

Our data indicate that treatment with atelizumab involves both cytolytic and antibody-mediated humoral effector mechanisms, respectively by T _eff And B cell mediation. This result was shown in the OAK study using a composite biomarker consisting of T effector 8-gene signature (tGE) and CD79A (OS HR less than the median relative to the median of the attlizumab arms) (HR =0.7, ci =0.55-0.91, p for the tGE in the attlizumab group relative to the less than median OS HR<0.01; for tGE + CD79A HR =0.65, ci =0.50-0.84, p<0.01 (FIGS. 13A to 13D) (Kowanetz, M. Et al, differential regulation of PD-L1 expression by immune and tumor cells in NSCLC and the response to treatment with atezolizumab (anti-PD-L1), proc. Natl. Acad. Sci. U.S. A115, E10119-E10126 (2018)). With respect to other alemtuzumab biomarkers, B cell characteristics correlated with an increase in PD-L1 on immune cells, likely reflecting an inflamed tumor microenvironment (fig. 10A to 10D). However, neither B-Cell nor TLS gene signature was associated with Tumor Mutation Burden (TMB) nor STK11 mutation (FIGS. 10A to 10D) (Gardara, D.R. et al, blood-based tumor mutation as a predictor of clinical findings in Non-Small-Cell Lung cancer peptides with interstitial colon tumor cells. Nat. Med.24,1441-1448 (2018); hellmann, M.D. et al, genomic Feature of Response Combination immunity in Patents with Advanced Non-Small-Lung cancer. Cancer Cell.33,843-852e (2018)).

Furthermore, our data show that the presence of B cells (especially plasma cells) within the tumor at baseline is a key factor in determining the OS benefit of NSCLC patients treated with atelizumab. To our knowledge, our data provide the first substantial evidence that the presence of plasma cells and the presence of organized lymphoid structures such as TLS at baseline are associated with astuzumab-mediated OS benefit, but not with chemotherapy-mediated OS benefit. To this end, transcriptomics data, including the largest public transcriptome NSCLC database (217 cases treated with CPI or chemotherapy and associated clinical outcomes), were analyzed according to two large independent randomized clinical trial cohorts (OAK, n =699; and POPLAR, n = 192).

In our study, gene expression analysis showed a strong correlation between B cell markers and OS benefit with atuzumab treatment. We used single cell analysis to derive and validate specific and robust gene signatures to reliably deconvolute follicular B cells, GC B cells and plasma cells from a large number of tumor transcriptomes. While each B cell subset has some correlation with the outcome, plasma cells appear to be the most important. This may indicate a productive local germinal center reaction in which mature plasma cells are the final product. These data are supported by similar significant clinical benefit for patients with histologically identified TLS or TLS-like structures. Taken together, these data indicate that in the case of tumors with high plasma cell infiltration, they are identified as tumors with TLS or TLS-like structures that, upon stimulation with CPI, provide sustained tumor control.

Interestingly, tumors with TLS or lymphoid aggregates accounted for approximately 30% of all NSCLC tumors analyzed and were rich in B/plasma cell characteristics. In our cohort, the presence of histologically-identified TLS was significantly associated with OS improvement following treatment with atuzumab, suggesting that they may play an important role in a sustained intratumoral immune response to PD- (L) 1 blockade. When we applied plasma cell characteristics to the TCGA NSCLC transcriptome that were not treated predominantly with CPI, it was found to be independent of overall survival, suggesting that plasma cell infiltration is not merely a prognosis for NSCLC. Overall, we show, in the context of two large randomized clinical trials against comparison of alemtuzumab to chemotherapy, that there is a strong predictive correlation between B/plasma cells and overall survival, which is unique to the mechanism of immunotherapy.

The mechanism by which B/plasma cells promote anti-tumor immunity is not known. Analysis showed that CD79+ B cells were in close proximity to CD8+ T cells when visualized by immunofluorescence imaging, providing support for the role of these cells as APCs. Class switching and memory B cells are enriched in tumors of CIT responders to melanoma and RCC, which may promote antigen presentation to major cytotoxic T cells.

Our analysis showed that there was a significant correlation between histologically defined TLS and B cell subset gene signatures, which clearly indicates a common underlying B cell biology, possibly including humoral immune responses contributing to the overall response to atuzumab. Our findings also clearly indicate that increased frequency of B and plasma cells at baseline correlates with improved clinical outcome in NSCLC patients treated with atuzumab, regardless of cytotoxic T cell characteristics.

In summary, our analysis shows that there is a significant correlation between histologically defined TLS, B cell IHC, TLS and germinal center-related gene signatures, indicating a common underlying B cell biology, including humoral immune responses contributing to the atlizumab-mediated response. Our findings also indicate that more B-cells and plasma cells were independent of T-cells at baseline, improving clinical outcomes in NSCLC patients treated with atelizumab.

Materials and methods

Patient population

The study was performed using tissue samples from open, randomized phase 2 POLAR (NCT 01903993) and phase 3 OAK test (NCT 02008227) which evaluated alemtuzumab with docetaxel (Rittmeyer, A. Et al, atezolizumab docetaxel in patients with clinical progress after receiving platinum chemotherapy, alpha phase 3, open-label, multicentranjd controlled clinical. Lance 389,255-265 (2017); fenthrenher, L. Et al, atezolizu olyzumab supplement for clinical course non-smooth-cell (POL) in NSCLC patients who progressed on platinum chemotherapy domised controlled batch. Lancet 387,1837-1846 (2016)). Tissue-derived RNA from Phase 2 FIR (NCT 01846416) and BIRCH (NCT 02031458) singles panel trials evaluated Abelmoschus mab (Peters, S. Et al Phase II Trial of Atzolizumab As First-Line or sublequent Therapy for Patients With Programmed Death-Ligand 1-Selected Advanced Non-Small-Cell-Lung Cancer (BIRCH). J.Clin. Oncol.35,2781-2789 (2017); spigel, D.R. et al, FIR: effect, saigy, and Biomarker of a Phase II Open-Label culture of PD-L1-L1731-Selected NSCLC.1748) in locally Advanced or metastatic NSCLCC Patients, were used, with reference. Patients in all trials received 1200mg of alemtuzumab IV (q 3 w) every 3 weeks until disease progression or loss of clinical benefit occurred, or 75mg/m ² Docetaxel IV q3w until PD occurs. Both FIR and BIRCH reported an improvement in response rate relative to historical averages; POPLAR and OAK studies showed that ATTETRAUzumab resulted in a significant improvement in OS compared to docetaxel regardless of PD-L1 expression (Rittmeyer, A. Et al, atezolizumab versatil docetaxel in tissues with previous linear displacement cam (OAK): a phase 3, open-lag, multicentre random coupled cam. Lance 389,255-265 (2017); fehrenger, L. Et al, atezolizumab versatil docetaxel for tissues with previous linear displacement cam-non-linear displacement cam-cell cam (POPLAR PLAR): multiple, open-lag, 2linear displacement cam 1846, 1836). All studies were approved by an independent ethics committee for each participating site, and safety data was reviewed by an independent data monitoring committee. No crossover is allowed and the primary endpoint is the OS.

Analysis of Gene expression

For 192 patients in POPLAR, 699 patients in OAK, 137 patients in FIR and 591 patients in BIRCH trial, all transcriptome profiles used the TruSeq RNA Access technique

And (4) generating. Ratio of RNAseq reads to ribosomal RNA sequencesAnd then to remove ribosomal reads. The NCI Build 38 human reference genome was then used to align the remaining reads of GSNAP version 2013-10-10, where at most two mismatches per 75 base sequence were allowed (parameter:' -M2-N10-B2-i 1-N1-w 200000-E1-pairmax-rna = 200000-clip-overlap). Transcript annotations were based on the Ensembl gene database (77 th edition). To quantify gene expression levels, the number of reads mapped to each RefSeq gene exon was calculated in a chain-specific manner using the functionality provided by the R Software package Genomic Alignments (Bioconductor) (Mariatasan, S. Et al, TGFbeta epitopes though responses to PD-L1 block by conditioning to exclusion of T cells. Nature 554,544-548 (2018); lawrence, M. Et al, software for computing and modeling Genomic sequences. PLoS. Comp. Biol.9, e1003118 (2013)).

To determine the biology associated with the benefits of alemtuzumab OS, patients with a survival period of <6 months (n = 24) or >12 months (n = 43) were grouped (Fehrenbacher, L. et al, atazolizumab versus facetaxel for patients with a previous treated non-small-cell lung cancer (POPLAR): a multicentre, open-label, phase 2 randulated controlled clinical. Lancet 387,1837-1846 (2016)). Differentially expressed genes between these two groups were determined using the R software package limma (Bioconductor) which estimates gene expression changes using a modest T-test using an empirical bayesian approach (Mariathasan, s. Et al, TGFbeta attributes pathway response to PD-L1 block by conditioning to expression of T cells. Nature 554,544-548 (2018); rietch, m.e. et al, limma powers differential expression analysis for RNA-sequencing and micro-array agents. Nucleic Acids res 43, e47 (2015).

Inter-dependence of key B-cell and T-cell gene contributions on overall survival benefit was also assessed using Cox proportional hazards regression analysis, among other things. In addition, paired t-test was used as indicated.

Genetic characterization

The B cell characteristics tested here include a number of markers and their various associated genes for naive B cells, memory B cells and plasma cells (Newman, a.m. et al, robust Expression of cell subsets from tissue Expression profiles. Nat. Methods 12,453-457 (2015); bindea, G.et al, spatiometric dynamics of endogenous animal cells derived from the animal in human cancer 39,782-795 (2013); iglesia, M.D. et al, genomic B-cell signatures using mRNA-seq in properties with sub-type-specific break and over-type cancer, res.20,3818-3829 (2014); palm, C.D., diehn, M.J., alizadeh, A.A. and Brown, P.O.cell-type specific Gene Expression profile of leucocytes in human biological module, BMC.Genomics7,115 (2006); suzuki, A. Et al, investigation of molecular biological Expression for diagnosis and analysis of environmental biological Expression (BMC, 12, expression of organism), M.D. et al, genomic B-cell signatures using mRNA-seq. No. 2. M.S. 2. C.. Initial B cell gene signature: ABCB4, BCL7A, BEND5, BRAF, IL4R, LINC00921, MEP1A, MICAL3, NIPSNAP3B, PSG2, SELL, TCL1A, UGT1A8, ZNF286A; memory B cell gene signature: AIM2, ALOX5, CLCA3P, FAM65B, IFNA10, IL7, NPIPB15, SP140, TNFRSF13B, TRAF4, ZBTB32; plasma cell gene characteristics: ABCB9, AMPD1, ANGPT4, ATXN8OS, C11, CCr10, HIST1H2AE, HIST1H2BG, IGHE, KCNA3, KCNG2, LOC100130100, MAN1A1, MANEA, MAST1, MROH7, MZB1, PAX7, PDK1, RASGRP3, REN, SPAG4, ST6GALNAC4, TGM5, UGT2B17, ZBP1, ZNF165; TLS Gene characterization as described in the review by Zhu et al (Zhu, G. Et al, tumor-Associated Tertiary lymphoma Structures: gene-Expression Profiling and the ir bioengineering. Front. Immunological.8, 767 (2017)): CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, CXCL13; b cell gene signature from this study: CD79A, SLAMF7, BTK, TNFRSF17 or BCMA, IGJ or JCHAIN, IGLL5, RBPJ, MZB1; and T cell effector gene characteristics: CD8A, EOMES, GZMA, TBX21, IFNG, GZMB, CXCL9, CXCL10 (Fehrenbacher, L. Et al, atezolizumab versals docetaxel for tasks with previous linear derived non-small-cell lung cancer (POPLAR): a multicentre, open-label, phase 2random controlled trial. Lancet 387,1837-1846 (2016)).

scRNAseq treatment and population specific characterization

Samples obtained from Gene Expression Omnibus (GEO) under accession number GSE131907 were counted as raw UMI per cell. In addition, the cell type and sample source annotation for each cell was also retrieved. These cell type annotations from the authors were used to isolate B cells from tumor and normal adjacent tissues and drain lymph nodes from the expression matrix. The Seurat software package (v3.1.4) was used for downstream analysis of B cells in R (3.6.2). Cells with more than 10% of fewer than 500 observed genes or mitochondrial reads were screened and the expression counts of the remaining cells were normalized to log (CPM/100 + 1). Principal component analysis was performed on the 2000 most variable genes and the first 30 principal components were used for UMAP dimensionality reduction and graph-based clustering. Clustering was determined using a resolution of 0.3. Markers for each cluster were detected by comparing all cells of the particular cluster to the rest of the cells in the dataset using Wilcoxon rank-sum test adjusted for multiple tests by Benjamini Hochberg method. non-B cell contaminating clusters (CD 3+ T cells, GZMB + LILRA4+ pDC, and HBA2+ erythrocytes) were removed from the initial dataset. After removal of these contaminating cells, the variable gene assay, PCA and UMAP dimensionality reduction were rerun using the parameters described above. Markers for each B cell subpopulation were identified as genes with adjusted p-values <0.001 and logFC >0.5, and B cells in the clusters were compared to all other cells in the dataset. To ensure B cell-specific expression of the marker, marker genes that are not expressed by B cells (including stromal cells, tumors, and non-B cell immune cells) in the complete data set (mean log (CPM/100 + 1) <1 in the non-B cell population) are further retained. Heatmaps for visual comparison were created using the pheatmap R software package (1.0.12).

CyTOF sample collection, staining and data processing

Six fresh NSCLC tumor samples were purchased from commercial suppliers (Discovery Life Sciences), which are part of adult patients undergoing surgical resection. After overnight fixation, cells were consumed in 3mL portions

Cell staining buffer washesAnd centrifuged at 800x g for 5 minutes. After aspiration of the wash buffer and resuspension of the cell pellet, 4mL was used

Water (Fluidigm) was used for another round of washing. Resuspend cells in 1mL

Water and counted. After obtaining a cell count, 3mL of the solution was added

Water, finally precipitating the cells once, and then performing instrumental collection. In the introduction of Helios ^TM Before, use

System (Fluidigm) resuspend pelleted cells in the medium containing EQ ^TM 1X of four element calibration bead (Fluidigm)

In water, filtration was then carried out using a 12X 75mm tube with a 35 μm nylon mesh cell filter cap (Corning). All FCS files are used

(MathWorks) normalization functions are normalized together and used

The software (FlowJo, LLC, ashland, oregon) performed the analysis. Protein marker expression intensities from mass cytometry analysis were aggregated from multiple samples and transformed using an inverse hyperbolic sine function. Applying dimensionality reduction to the transformed expression matrix using a Unified Manifold Approximation and Projection (UMAP) package with the following default parameters: min _ dist:0.1, n _ neighbors:15, n _ components:2, and metric: euclidean. In both the CD45+ and CD8+ populations, a single sample was downsampled to 8,000 cells per sample. Attach UMAP coordinates behind the ". Fcs" file as hands for use in connection with FlowJo (LLC) Dynamic gating analysis integrated additional channels.

Immunofluorescence and tertiary lymphoid analysis

Triple immunofluorescence (CD 8/CD 79/Ki-67) analysis was performed on 4 μm sections of Formalin Fixed Paraffin Embedded (FFPE) tumor samples, followed by deparaffinization, rehydration and epitope repair with antigen repair solution pH 6 at 99 ℃ for 20 minutes. Each round of staining began with quenching in 3% hydrogen peroxide. Sections were incubated in anti-CD 79a rabbit monoclonal antibody (SP 18) (Thermo Fischer Scientific, MA 5-14556) diluted in 1. Elution was carried out in antigen retrieval solution preheated to 99 ℃ for 20 minutes. Sections were incubated in anti-CD 8 mouse monoclonal antibody C8/144B (Dako, M7103) diluted to 1.5. Mu.g/ml, detected with PowerVision polyHRP anti-mouse antibody, and amplified with Alexa-Fluor 647 tyramide. After the elution step, sections were incubated in anti-Ki 67 mouse monoclonal antibody (MIB-1) (Dako, M7240) diluted to 1 μ g/ml, detected with PowerVision poly-HRP anti-mouse antibody, and amplified with Alexa-Fluor 555 tyramide. Slides were counterstained with 4', 6-diamidino-2-phenylindole (DAPI). Images of the fluorescent slides were collected on a NanoZoomer XR.

TLS was identified using H & E stained FFPE sections and examined by a pathologist (HK) to identify lymphatic aggregates similar to TLS. TLS is defined as those regions with well-defined boundaries that are morphologically similar to lymph aggregates (Dieu-Nosjean, M.C. et al, tele-physiology structures in cancer and beyond, trends immunol.35,571-580 (2014)). Representative slides were selected for IHC using MECA-79 in combination with peripheral lymph node addressin.

BCR lineage analysis

Of the 32 tumor samples, 16 (9 from BIRCH and 7 from FIR) were collected from NSCLC patients receiving treatment with Attemplumab (Tan, P. Et al, regulatory role of the CXCL13-CXCR5 axis in the tumor micro environmental, precision in clin. Med.1,49-56 (2018); bruno T.C.evaluating the anticancer role of the B cells in tissues with non-small cell lung cancer. J.clin. Oncol.35,75 (2017)). RNA was extracted from FFPE tumors using a high purity FFPE RNA isolation kit (roche). The BCR cDNA library was generated using the Immunoverse IgH kit (ArcherDX, inc., boulder, CO). The library was quantified using a KAPA Universal library quantification kit (KAPA Biosystems, wilmington, mass.). The libraries were combined at equimolar concentrations and sequenced using MiSeq kit v3 (600 cycles) and Illumina MiSeq instrument (Illumina, inc. By using

The analysis software analyzed the sequence data to calculate the SDI for each library (Greiff, V. et al, A bioinformatic frame for immune response sensitivity development of immune status. Genome Med.7,49 (2015); kaplinsky, J. et al, road tests of immune response sensitivity from high-throughput memories on samples. Nat. Commun.7,11881 (2016)). Statistical analysis of SDI between pairs of samples from the same patient was performed using Wilcoxon paired signed rank sum test and variation in SDI before/after treatment was compared between different patient groups using the Mann Whitney test.

Other examples

Although the foregoing invention has been described in some detail by way of illustration and embodiments for purposes of clarity of understanding, these descriptions and embodiments should not be construed as limiting the scope of the invention. The disclosures of all patent and scientific literature cited herein are expressly incorporated by reference in their entirety.

Exemplary embodiments of the present invention include those enumerated below.

1. A method of identifying an individual having cancer who is likely to benefit from treatment with a binding antagonist comprising a PD-L1 axis, the method comprising determining the expression levels of two or more of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ and MZB1 in a sample from the individual, wherein the individual is identified as an individual who is likely to benefit from treatment with a binding antagonist comprising a PD-L1 axis if the immune score expression levels of the two or more genes are greater than the reference immune score expression levels of the two or more genes.

2. A method of selecting a therapy for an individual having cancer, the method comprising determining the expression levels of two or more of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ and MZB1 in a sample from the individual, wherein the individual is identified as an individual likely to benefit from treatment comprising a PD-L1 axis binding antagonist if the immune score expression levels of the two or more genes are greater than the reference immune score expression levels of the two or more genes.

3. The method of example 1 or 2, wherein the immune score expression level of two or more genes in the sample is greater than the reference immune score expression level, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist.

4. A method of treating an individual having cancer, the method comprising:

(a) Determining the expression level of two or more of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein the determination that the immune score expression level of the two or more genes in the sample is higher than the reference immune score expression level of the two or more genes; and

5. A method of treating cancer in an individual, wherein the individual has been determined to have in a sample from the individual an immune score expression level for two or more of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ and MZB1 that is greater than a reference immune score expression level for the two or more genes, the method comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist.

6. The method of any one of embodiments 1 to 5, wherein the immune score reference expression level is an immune score expression level of two or more genes in a reference population.

7. The method of embodiment 6, wherein the reference population is a population of individuals having cancer.

8. The method of embodiment 7, wherein the population of individuals comprises a first subset of individuals who have received treatment with a PD-L1 axis binding antagonist and a second subset of individuals who have received treatment with a therapy that does not contain a PD-L1 axis binding antagonist.

9. The method of embodiment 8, wherein the reference immune score expression level visibly differentiates each of the first subset of individuals from the second subset of individuals based on a significant difference between the individual's responsiveness to treatment with the PD-L1 axis binding antagonist and the individual's responsiveness to treatment with the therapy without the PD-L1 axis binding antagonist that is greater than the reference immune score expression level, wherein the individual's responsiveness to treatment with the PD-L1 axis binding antagonist is significantly improved relative to the individual's responsiveness to treatment with the therapy without the PD-L1 axis binding antagonist.

10. The method of

embodiment

8 or 9, wherein the therapy without the PD-L1 axis binding antagonist comprises an anti-tumor agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, or a combination thereof.

11. The method of embodiment 10, wherein the therapy without the PD-L1 axis binding antagonist comprises a chemotherapeutic agent.

12. The method of embodiment 11, wherein the chemotherapeutic agent is docetaxel.

13. The method of any one of embodiments 9 to 12, wherein responsiveness to treatment comprises prolongation of OS, prolongation of Progression Free Survival (PFS), or an increase in determined best overall response (BCOR).

14. The method of embodiment 13, wherein responsiveness to treatment comprises prolongation of OS.

15. The method of any one of embodiments 6-14, wherein the reference immune score expression level is the median of the expression levels of each of the two or more genes in the reference population.

16. The method of any one of embodiments 1-15, wherein genes comprise three or more of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB 1.

17. The method of any one of embodiments 1-16, wherein genes comprise four or more of CD79A, CD19, BANK1, jchan, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB 1.

18. The method of any one of embodiments 1-17, wherein genes comprise five or more of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1.

19. The method of any one of embodiments 1-18, wherein genes comprise six or more of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1.

20. The method of any one of embodiments 1-19, wherein genes comprise seven, eight, nine, ten or more of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1.

21. The method of any one of embodiments 1-20, wherein genes comprise CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1.

22. A method of identifying an individual having cancer who is likely to benefit from treatment with a PD-L1 axis binding antagonist, the method comprising determining the expression level of one or more of the genes CD79A, CD19, BANK1, jchanin, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein the benefit comprises an increase in the overall phase (OS) of the individual as compared to treatment without the PD-L1 axis binding antagonist if the immune score expression level of the one or more genes is greater than a reference immune score expression level for the one or more genes.

23. A method of selecting a therapy for an individual having cancer, the method comprising determining the expression level of one or more of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein in the event that the immune score expression level of the one or more genes is greater than a reference immune score expression level for the one or more genes, identifying the individual as an individual who is likely to benefit from treatment with a PD-L1 axis binding antagonist, wherein the benefit comprises an increase in OS in the individual as compared to treatment without the PD-L1 axis binding antagonist.

24. The method of

embodiment

22 or 23, wherein the immune score expression level of one or more genes in the sample is greater than the reference immune score expression level, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist.

25. A method of treating an individual having cancer, the method comprising:

(a) Determining the expression level of one or more of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein determining the immune score expression level of the one or more genes in the sample is greater than a reference immune score expression level for the one or more genes identifies the individual as likely to benefit from treatment with a PD-L1 axis binding antagonist, wherein the benefit comprises a prolongation of OS in the individual as compared to treatment without the PD-L1 axis binding antagonist; and

26. A method of treating cancer in an individual, wherein the individual has been determined to have an immune score expression level of one or more of the genes CD79A, CD19, BANK1, jchan, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual that is greater than a reference immune score expression level of the one or more genes, the method comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist, wherein in the event the immune score expression level of the one or more genes is greater than the reference immune score expression level of the one or more genes, identifying the individual as an individual likely to benefit from treatment comprising a PD-L1 axis binding antagonist, wherein the benefit comprises an extended OS in the individual as compared to treatment without the PD-L1 axis binding antagonist.

27. The method of any one of embodiments 22-26, wherein the immune score expression level of one of genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 is determined.

28. The method of embodiment 27, wherein the immune score expression level of CD79A is determined.

29. The method of any one of embodiments 22 to 28, wherein the immune score reference expression level is an immune score expression level of one or more genes in a reference population.

30. The method of embodiment 29, wherein the reference population is a population of individuals with cancer.

31. The method of embodiment 30, wherein the population of individuals comprises a first subset of individuals who have received treatment with a PD-L1 axis binding antagonist and a second subset of individuals who have received treatment with a therapy that does not contain a PD-L1 axis binding antagonist.

32. The method of embodiment 31, wherein the immune score reference expression level visibly distinguishes between the first subset of individuals and each of the second subset of individuals based on a significant difference between the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist and the responsiveness of the individual to treatment with the therapy without the PD-L1 axis binding antagonist that is greater than the reference immune score expression level, wherein the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist is significantly improved relative to the responsiveness of the individual to treatment with the therapy without the PD-L1 axis binding antagonist.

33. The method of

embodiment

31 or 32, wherein the therapy without the PD-L1 axis binding antagonist comprises an anti-neoplastic agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, or a combination thereof.

34. The method of embodiment 33, wherein the therapy without the PD-L1 axis binding antagonist comprises a chemotherapeutic agent.

35. The method of embodiment 34, wherein the chemotherapeutic agent is docetaxel.

36. The method of any one of embodiments 32-35, wherein responsiveness to treatment comprises prolongation of OS, prolongation of PFS, or an increase in BCOR.

37. The method of embodiment 36, wherein responsiveness to treatment is an extension of Overall Survival (OS).

38. The method of any one of embodiments 29 to 37, wherein the immune score reference expression level is the median of the expression levels of each of the one or more genes in the reference population.

39. The method of embodiment 38, wherein the median expression level is the median of the average Z-scores of the expression levels of each of the two or more genes in the reference population.

40. The method of any one of embodiments 22-39, wherein genes comprise two or more of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB 1.

41. The method of any one of embodiments 1-5 and 40, wherein the two or more genes comprise TNFRSF17 and IGJ.

42. The method according to any one of embodiments 1-5 and 41, wherein the two genes consist of TNFRSF17 and IGJ.

43. The method of any one of embodiments 22-41, wherein genes comprise three or more of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB 1.

44. The method of any one of embodiments 22-41-43, wherein genes comprise four or more of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB 1.

45. The method of any one of embodiments 22-41, 43 and 44, wherein the genes comprise five or more of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB 1.

46. The method of any one of embodiments 22-41 and embodiments 43-45, wherein genes comprise six or more of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB 1.

47. The method of any one of embodiments 22-41 and embodiments 43-46, wherein the genes comprise seven, eight, nine, ten or more of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB 1.

48. The method of any one of embodiments 22-41 and 43-47, wherein the genes comprise CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1.

49. The method of embodiment 48, wherein the gene consists of CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1.

50. A method of identifying an individual having cancer who is likely to benefit from treatment with a PD-L1 axis binding antagonist, the method comprising determining the expression level of one or more of the genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7 and IGLL5 in a sample from the individual, wherein the individual is identified as an individual who is likely to benefit from treatment with a PD-L1 axis binding antagonist if the immune score expression level of the one or more genes is higher than a reference immune score expression level of the one or more genes.

51. A method of selecting a therapy for an individual having cancer, the method comprising determining the expression level of one or more of the genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7 and IGLL5 in a sample from the individual, wherein the individual is identified as an individual likely to benefit from treatment comprising a PD-L1 axis binding antagonist if the immune score expression level of the one or more genes is higher than the reference immune score expression level of the one or more genes.

52. The method of

embodiment

50 or 51, wherein the immune score expression level of one or more genes in the sample is greater than the reference immune score expression level, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist.

53. A method of treating a subject having cancer, the method comprising:

54. A method of treating cancer in an individual, wherein the individual has been determined to have an immune score expression level of one or more of the genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from the individual that is higher than a reference immune score expression level for the one or more genes, the method comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist.

55. The method of any one of embodiments 50 to 54, wherein the immune score reference expression level is the immune score expression level of one or more genes in a reference population.

56. The method of embodiment 55, wherein the reference population is a population of individuals with cancer.

57. The method of embodiment 56, wherein the population of individuals comprises a first subset of individuals who have received treatment with a PD-L1 axis binding antagonist and a second subset of individuals who have received treatment with a therapy that does not contain a PD-L1 axis binding antagonist.

58. The method of embodiment 57, wherein the reference immune score expression level visibly differentiates each of the first subset of individuals from the second subset of individuals based on a significant difference between the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist and the responsiveness of the individual to treatment with the therapy without the PD-L1 axis binding antagonist that is greater than the reference immune score expression level, wherein the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist is significantly improved relative to the responsiveness of the individual to treatment with the therapy without the PD-L1 axis binding antagonist.

59. The method of

embodiment

57 or 58, wherein the therapy without the PD-L1 axis binding antagonist comprises an anti-neoplastic agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, or a combination thereof.

60. The method of embodiment 59, wherein the therapy without the PD-L1 axis binding antagonist comprises a chemotherapeutic agent.

61. The method of embodiment 60, wherein the chemotherapeutic agent is docetaxel.

62. The method of any one of embodiments 58 to 61, wherein responsiveness to treatment comprises prolongation of OS, prolongation of PFS, or an increase in BCOR.

63. The method of embodiment 62, wherein responsiveness to treatment is an extension of Overall Survival (OS).

64. The method of any one of embodiments 55 to 63, wherein the reference immune score expression level is the median of the expression levels of each of the one or more genes in the reference population.

65. The method of any one of embodiments 50-64, wherein genes comprise two or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

66. The method of any one of embodiments 50-65, wherein genes comprise three or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

67. The method of any one of embodiments 50-66, wherein genes comprise four or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

68. The method of any one of embodiments 50-67, wherein genes comprise five or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

69. The method of any one of embodiments 50-68, wherein genes comprise six or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

70. The method of any one of embodiments 50-69, wherein genes comprise seven or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

71. The method of any one of embodiments 50-70, wherein genes comprise eight or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

72. The method of any one of embodiments 50-71, wherein the genes comprise nine or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

73. The method of any one of embodiments 50-72, wherein the genes comprise 10 or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

74. The method of any one of embodiments 50-73, wherein genes comprise 11 or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

75. The method of any one of embodiments 50-74, wherein genes comprise 12 or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

76. The method of any one of embodiments 50-75, wherein genes comprise 13 or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

77. The method of any one of embodiments 50-76, wherein genes comprise MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5.

78. A method of identifying an individual having cancer who is likely to benefit from treatment with a PD-L1 axis binding antagonist, the method comprising determining the expression level of one or more of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7 and IGLL5 in a sample from the individual, wherein the individual is identified as an individual who is likely to benefit from treatment with the PD-L1 axis binding antagonist if the immune score expression level of the one or more genes is greater than a reference immune score expression level for the one or more genes, wherein the benefit comprises an extension of the OS of the individual as compared to treatment without the PD-L1 axis binding antagonist.

79. A method of selecting a therapy for an individual having cancer, the method comprising determining the expression level of one or more of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from the individual, wherein the individual is identified as an individual likely to comprise a treatment with a PD-L1 axis binding antagonist if the immune score expression level of the one or more genes is greater than a reference immune score expression level for the one or more genes, wherein the benefit comprises a prolonged OS for the individual as compared to treatment without the PD-L1 axis binding antagonist.

80. The method of

embodiment

78 or 79, wherein the immune score expression level of one or more genes in the sample is greater than the reference immune score expression level, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist.

81. A method of treating an individual having cancer, the method comprising:

82. A method of treating cancer in an individual, wherein the individual has been determined to have an immune score expression level of one or more of the genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7 and IGLL5 in a sample from the individual that is higher than a reference immune score expression level for the one or more genes, the method comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist, wherein the individual is identified as an individual who is likely to comprise treatment with the PD-L1 axis binding antagonist, wherein the benefit comprises an extended OS in the individual as compared to treatment without the PD-L1 axis binding antagonist.

83. The method of any one of embodiments 78 to 82, wherein an immune score reference expression level is an immune score expression level of one or more genes in a reference population.

84. The method of embodiment 83, wherein the reference population is a population of individuals with cancer.

85. The method of embodiment 84, wherein the population of individuals comprises a first subset of individuals who have received treatment with a PD-L1 axis binding antagonist and a second subset of individuals who have received treatment with a therapy that does not contain a PD-L1 axis binding antagonist.

86. The method of embodiment 85, wherein the immune score reference expression level visibly distinguishes between the first subset of individuals and each of the second subset of individuals based on a significant difference between the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist and the responsiveness of the individual to treatment with the therapy without the PD-L1 axis binding antagonist that is greater than the reference immune score expression level, wherein the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist is significantly improved relative to the responsiveness of the individual to treatment with the therapy without the PD-L1 axis binding antagonist.

87. The method of

embodiment

85 or 86, wherein the therapy without the PD-L1 axis binding antagonist comprises an anti-tumor agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, or a combination thereof.

88. The method of embodiment 87, wherein the therapy without the PD-L1 axis binding antagonist comprises a chemotherapeutic agent.

89. The method of embodiment 88, wherein the chemotherapeutic agent is docetaxel.

90. The method of any one of embodiments 86-89, wherein responsiveness to treatment comprises prolongation of OS, prolongation of PFS, or an increase in BCOR.

91. The method of embodiment 90, wherein responsiveness to treatment is an extension of Overall Survival (OS).

92. The method of any one of embodiments 83-91, wherein the immune score reference expression level is the median of the expression levels of each of the one or more genes in the reference population.

93. The method of embodiment 92, wherein the median expression level is the median of the average Z scores for the expression levels of each of the one or more genes in the reference population.

94. The method of any one of embodiments 78 to 93, wherein genes comprise two or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

95. The method of any one of embodiments 78 to 94, wherein genes comprise three or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

96. The method of any one of embodiments 78 to 95, wherein genes comprise four or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

97. The method of any one of embodiments 78 to 96, wherein genes comprise five or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

98. The method of any one of embodiments 78 to 97, wherein genes comprise six or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

99. The method of any one of embodiments 78 to 98, wherein genes comprise seven or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

100. The method of any one of embodiments 78 to 99, wherein genes comprise eight or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

101. The method of any one of embodiments 78 to 100, wherein genes comprise nine or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

102. The method of any one of embodiments 78 to 101, wherein genes comprise 10 or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

103. The method of any one of embodiments 78 to 102, wherein genes comprise 11 or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

104. The method of any one of embodiments 78 to 103, wherein genes comprise 12 or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

105. The method of any one of embodiments 78 to 104, wherein genes comprise 13 or more of MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL 5.

106. The method of any one of embodiments 78 to 105, wherein genes comprise MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5.

107. A method of identifying an individual having cancer who is likely to benefit from treatment comprising a PD-L1 axis binding antagonist, the method comprising determining the presence of Tertiary Lymphoid Structure (TLS) in a tumor sample from the individual, wherein the presence of TLS in the tumor sample identifies the individual as an individual who is likely to benefit from treatment comprising a PD-L1 axis binding antagonist.

108. A method of selecting a therapy for an individual having cancer, the method comprising determining the presence of TLS in a tumor sample from the individual, wherein the presence of TLS in the tumor sample identifies the individual as an individual who is likely to benefit from treatment comprising a PD-L1 axis binding antagonist.

109. The method of

embodiment

107 or 108, wherein the presence of TLS is determined in a sample from the individual, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist.

110. A method of treating an individual having cancer, the method comprising:

(a) Determining the presence of TLS in a tumor sample from the individual; and

111. A method of treating cancer in an individual, wherein the individual has been determined to have TLS present in a tumor sample from the individual, comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist.

112. The method of any one of embodiments 107 to 111, wherein the presence of TLS is determined by histological staining, immunohistochemistry (IHC), immunofluorescence, or gene expression analysis.

113. The method of embodiment 112, wherein the histological staining comprises hematoxylin and eosin (H & E) staining.

114. The method of embodiment 112, wherein IHC or immunofluorescence comprises detecting CD62L, L-selectin, CD40 or CD8.

115. The method of embodiment 114, wherein the CD62L or L-selectin is detected using a MECA-79 antibody.

116. The method of embodiment 112, wherein the gene expression analysis comprises determining the expression level of a TLS gene signature in the sample.

117. The method of embodiment 116, wherein a TLS gene characteristic comprises one or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13.

118. The method of embodiment 117, wherein the genes comprise two or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13.

119. A method of identifying an individual having cancer who is likely to benefit from treatment comprising a PD-L1 axis binding antagonist, the method comprising determining the expression level of two or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual, wherein the individual is identified as an individual who is likely to benefit from treatment comprising a PD-L1 axis binding antagonist if the immune score expression level of the two or more genes is greater than a reference immune score expression level of the two or more genes.

120. A method of selecting a therapy for an individual having a cancer, the method comprising determining the expression level of two or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual, wherein the individual is identified as an individual likely to benefit from treatment comprising a PD-L1 axis binding antagonist if the immune score expression level of the two or more genes is greater than a reference immune score expression level of the two or more genes.

121. The method of embodiment 119 or 120, wherein the immune score expression level of five or more genes in the sample is greater than the reference immune score expression level, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist.

122. A method of treating an individual having cancer, the method comprising:

(a) Determining the expression level of two or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual, wherein determining the immune score expression level of the two or more genes in the sample is higher than a reference immune score expression level of the two or more genes; and

123. A method of treating cancer in an individual, wherein the individual has been determined to have an immune score expression level of two or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual that is higher than a reference immune score expression level of the two or more genes, the method comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist.

124. The method of any one of embodiments 119-123, wherein the reference immune score expression level is an immune score expression level of two or more genes in a reference population.

125. The method of embodiment 124, wherein the reference population is a population of individuals with cancer.

126. The method of embodiment 125, wherein the population of individuals comprises a first subset of individuals who have received treatment with a PD-L1 axis binding antagonist and a second subset of individuals who have received treatment with a therapy that does not contain a PD-L1 axis binding antagonist.

127. The method of embodiment 126, wherein the reference immune score expression level visibly distinguishes each of the first subset of individuals from the second subset of individuals based on a significant difference between the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist and the responsiveness of the individual to treatment with the therapy without the PD-L1 axis binding antagonist above the reference expression level, wherein the responsiveness of the individual to treatment with the PD-L1 axis binding antagonist is significantly improved relative to the responsiveness of the individual to treatment with the therapy without the PD-L1 axis binding antagonist.

128. The method of embodiment 126 or 127, wherein the therapy without a PD-L1 axis binding antagonist comprises an anti-tumor agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, or a combination thereof.

129. The method of embodiment 128, wherein the therapy without the PD-L1 axis binding antagonist comprises a chemotherapeutic agent.

130. The method of embodiment 129, wherein the chemotherapeutic agent is docetaxel.

131. The method of any one of embodiments 127 to 130, wherein responsiveness to treatment comprises prolongation of OS, prolongation of PFS, or an increase in BCOR.

132. The method of embodiment 131, wherein responsiveness to treatment is an extension of Overall Survival (OS).

133. The method of any one of embodiments 125-132, wherein the reference immune score expression level is the median of the expression levels of each of the two or more genes in the reference population.

134. The method of embodiment 133, wherein the median expression level is the median of the average Z-scores of the expression levels of each of the two or more genes in the reference population.

135. The method of any one of embodiments 110-134, wherein the genes comprise three or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13.

136. The method of any one of embodiments 119-135, wherein a gene comprises four or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13.

137. The method of any one of embodiments 119-136, wherein a gene comprises five or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13.

138. The method of any one of embodiments 119-137, wherein the genes comprise six or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13.

139. The method of any one of embodiments 119-138, wherein genes comprise seven or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13.

140. The method of any one of embodiments 119-139, wherein genes comprise eight or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13.

141. The method of any one of embodiments 119-140, wherein genes comprise nine or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13.

142. The method of any one of embodiments 119-141, wherein genes comprise ten or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL 13.

143. The method of any one of embodiments 119-142, wherein a gene comprises eleven or more of CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13.

144. The method of any one of embodiments 119-143, wherein the gene comprises CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13.

145. The method of any one of embodiments 1-106 and 119-144, wherein the expression level is a nucleic acid expression level.

146. The method of embodiment 145, wherein the nucleic acid expression level is an mRNA expression level.

147. The method of embodiment 146, wherein mRNA expression levels are determined by RNA-seq, RT-qPCR, multiplex qPCR or RT-qPCR, microarray analysis, SAGE, massARRAY technology, FISH, or a combination thereof.

148. The method of embodiment 147, wherein mRNA expression levels are detected using RNA-seq.

149. The method of any one of embodiments 1 to 106 and 119 to 144, wherein the expression level is a protein expression level.

150. The method of embodiment 149, wherein the protein expression level is determined by IHC, immunofluorescence, mass spectrometry, flow cytometry, and western blot, or a combination thereof.

151. The method of any one of embodiments 1-106 and 119-150, wherein the expression level is detected in tumor cells, tumor infiltrating immune cells, stromal cells, paracancerous normal tissue (NAT) cells, or a combination thereof.

152. A method of identifying an individual having cancer who is likely to benefit from treatment with a binding antagonist comprising PD-L1 axis, the method comprising determining the number of B cells in a tumor sample from the individual, wherein in the event that the number of B cells in the tumor sample is greater than a reference number of B cells, identifying the individual as an individual who is likely to benefit from treatment with a binding antagonist comprising PD-L1 axis.

153. A method of selecting a therapy for an individual having cancer, the method comprising determining the number of B cells in a tumor sample from the individual, wherein in the event the number of B cells in the tumor sample is greater than a reference number of B cells, identifying the individual as an individual likely to benefit from treatment comprising a PD-L1 axis binding antagonist.

154. The method of

embodiment

152 or 153, wherein the number of B cells in the sample is greater than the reference number, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist.

155. A method of treating an individual having cancer, the method comprising:

(a) Determining the number of B cells in a tumor sample from an individual; and

156. A method of treating cancer in an individual, wherein the individual has been determined to have a number of B cells in a tumor sample from the individual that is greater than a reference number of B cells, comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist.

157. The method of any one of embodiments 152-156, wherein B cells comprise CD79+ B cells, igG + B cells, and/or plasma cells.

158. The method of embodiment 157, wherein the B cells comprise plasma cells.

159. A method of identifying an individual having cancer who is likely to benefit from treatment comprising a PD-L1 axis binding antagonist, the method comprising determining whether the individual has clonally expanded B cells in a tumor sample from the individual, wherein the clonally expanded B cells in the sample identify the individual as an individual who is likely to benefit from treatment comprising a PD-L1 axis binding antagonist.

160. A method of selecting a therapy for an individual having cancer, the method comprising determining whether the individual has clonally expanded B cells in a tumor sample from the individual, wherein the clonally expanded B cells in the sample identify the individual as an individual likely to benefit from treatment comprising a PD-L1 axis binding antagonist.

161. The method of

embodiment

159 or 160, wherein the tumor sample comprises clonally expanded B cells, and the method further comprises administering to the individual an effective amount of a PD-L1 axis binding antagonist.

162. A method of treating a subject having cancer, the method comprising:

(a) Determining that the individual has clonally expanded B cells in a tumor sample from the individual; and

163. A method of treating cancer in an individual, wherein the individual has been determined to have clonally expanded B cells in a tumor sample from the individual, comprising administering to the individual an effective amount of a PD-L1 axis binding antagonist.

164. The method of any one of embodiments 159 to 163, wherein the clonally expanded B cells are clonally expanded plasma cells.

165. The method of any one of embodiments 159 to 164, wherein clonally expanded B cells are detected by measuring the diversity of B Cell Receptor (BCR) gene lineages in a tumor sample.

166. The method of embodiment 165, wherein in the event that a Shannon Diversity Index (SDI) of a BCR gene lineage in a tumor sample from the individual is lower than a reference SDI, identifying the individual as an individual likely to benefit from treatment comprising a PD-L1 axis binding antagonist.

167. The method of any one of embodiments 1-106 and embodiments 119-151, wherein the sample is a tissue sample, a cell sample, a whole blood sample, a plasma sample, a serum sample, or a combination thereof.

168. The method of embodiment 167, wherein the tissue sample is a tumor tissue sample.

169. The method of any one of embodiments 107-118 and embodiments 152-166, wherein the tumor sample is a tumor tissue sample.

170. The method of embodiment 168 or 169, wherein the tumor tissue sample comprises tumor cells, tumor infiltrating immune cells, stromal cells, NAT cells, or a combination thereof.

171. The method of any one of embodiments 168 to 170, wherein the tumor tissue sample is a Formalin Fixed and Paraffin Embedded (FFPE) sample, an archived sample, a fresh sample, or a frozen sample.

172. The method of embodiment 171, wherein the tumor tissue sample is an FFPE sample.

173. The method of any one of embodiments 1-172, wherein cancer is lung cancer, kidney cancer, bladder cancer, breast cancer, colorectal cancer, ovarian cancer, pancreatic cancer, stomach cancer, esophageal cancer, mesothelioma, melanoma, head and neck cancer, thyroid cancer, sarcoma, prostate cancer, glioblastoma, cervical cancer, thymus cancer, leukemia, lymphoma, myeloma, mycosis fungoides, merkel cell carcinoma, or hematological malignancy.

174. The method of embodiment 173, wherein the cancer is lung, kidney, bladder or breast cancer.

175. The method of embodiment 174, wherein the lung cancer is non-small cell lung cancer (NSCLC).

176. The method of embodiment 175, wherein the NSCLC is non-squamous NSCLC.

177. The method of embodiment 175, wherein the NSCLC is squamous NSCLC.

178. The method according to any one of examples 1-3, examples 6-52, examples 55-109, examples 112-121, examples 124-154, examples 157-161, and examples 164-177, wherein the benefit comprises prolongation of OS in the individual, prolongation of PFS in the individual, and/or improvement in BCOR in the individual as compared to treatment without the PD-L1 axis binding antagonist.

179. The method of embodiment 178, wherein the benefit comprises a prolongation of OS in the individual as compared to treatment without the PD-L1 axis binding antagonist.

180. The method of any one of embodiments 1 to 179, wherein the PD-L1 axis binding antagonist is selected from the group consisting of a PD-L1 binding antagonist, a PD-1 binding antagonist, and a PD-L2 binding antagonist.

181. The method of embodiment 180, wherein the PD-L1 axis binding antagonist is a PD-L1 binding antagonist.

182. The method of embodiment 181, wherein the PD-L1 binding antagonist inhibits the binding of PD-L1 to one or more of its ligand binding partners.

183. The method of embodiment 182, wherein the PD-L1 binding antagonist inhibits the binding of PD-L1 to PD-1.

184. The method of embodiment 182, wherein the PD-L1 binding antagonist inhibits the binding of PD-L1 to B7-1.

185. The method of any one of embodiments 182 to 184, wherein the PD-L1 binding antagonist inhibits the binding of PD-L1 to both PD-1 and B7-1.

186. The method of any one of embodiments 182 to 185, wherein the PD-L1 binding antagonist is an antibody or antigen-binding fragment thereof.

187. The method of embodiment 186, wherein the antibody is selected from the group consisting of: alemtuzumab, MDX-1105, MEDI4736 (devoluumab), and MSB0010718C (avizumab).

188. The method of embodiment 186, wherein the antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

189. The method of embodiment 186, wherein the antibody comprises: a heavy chain variable region comprising the amino acid sequence of SEQ ID NO 7; and a light chain variable region comprising the amino acid sequence of SEQ ID NO 8.

190. The method of embodiment 180, wherein the PD-L1 axis binding antagonist is a PD-1 binding antagonist.

191. The method of embodiment 190, wherein the PD-1 binding antagonist inhibits the binding of PD-1 to one or more of its ligand binding partners.

192. The method of embodiment 191, wherein the PD-1 binding antagonist inhibits the binding of PD-1 to PD-L1.

193. The method of embodiment 191, wherein the PD-1 binding antagonist inhibits the binding of PD-1 to PD-L2.

194. The method of any one of embodiments 191-193, wherein the PD-1 binding antagonist inhibits the binding of PD-1 to both PD-L1 and PD-L2.

195. The method of any one of embodiments 191 to 194, wherein the PD-1 binding antagonist is an antibody or antigen-binding fragment thereof.

196. The method of embodiment 195, wherein the antibody is selected from the group consisting of: MDX-1106 (nivolumab), MK-3475 (pembrolizumab), MEDI-0680 (AMP-514), PDR001, REGN2810, and BGB-108.

197. The method of any one of embodiments 190 to 194, wherein the PD-1 binding antagonist is an Fc fusion protein.

198. The method of embodiment 197, wherein the Fc fusion protein is AMP-224.

199. The method of any one of embodiments 1-198, wherein the subject has not been treated for the cancer.

200. The method of embodiment 199, wherein the individual has not been administered a PD-L1 axis binding antagonist.

201. The method of

embodiment

199 or 200, wherein the cancer is NSCLC and the individual does not have EGFR or ALK genomic tumor aberrations.

202. The method of any one of embodiments 1-198, wherein the subject has been previously treated for the cancer.

203. The method of embodiment 202, wherein the individual has been treated for the cancer by administering to the individual a platinum-containing chemotherapeutic, and wherein the individual is non-responsive to the chemotherapeutic.

204. The method according to any one of embodiments 4 to 21, embodiments 25 to 48, embodiments 53 to 77, embodiments 81 to 106, embodiments 110 to 118, embodiments 122 to 151, embodiments 155 to 158 and embodiments 162 to 203, wherein the PD-L1 axis binding antagonist is administered as a monotherapy.

205. The method according to any one of embodiments 4 to 21, embodiments 25 to 48, embodiments 53 to 77, embodiments 81 to 106, embodiments 110 to 118, embodiments 122 to 151, embodiments 155 to 158 and embodiments 162 to 203, wherein the method further comprises administering an effective amount of one or more additional therapeutic agents.

206. The method of embodiment 205, wherein the one or more additional therapeutic agents comprise an anti-neoplastic agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, an immunomodulatory agent, or a combination thereof.

207. The method of any one of embodiments 1-206, wherein the subject is a human.

208. A kit comprising a PD-L1 axis binding antagonist and instructions for administering the PD-L1 axis binding antagonist to an individual who has been identified as an individual who is likely to benefit from treatment comprising the PD-L1 binding antagonist according to the method of any one of examples 1 to 3, examples 6 to 52, examples 55 to 109, examples 111 to 121, examples 124 to 154, examples 157 to 161, and examples 164 to 203.

209. A kit comprising a PD-L1 axis binding antagonist and instructions for administering the PD-L1 axis binding antagonist to an individual for whom treatment comprising the PD-L1 axis binding antagonist has been selected according to the method of any one of example 2, example 3, examples 6-21, example 23, examples 27-49, example 51, examples 55-77, example 79, examples 83-106, example 108, examples 112-118, example 120, examples 124-151, example 153, 154, 157, 158, 160, 161, and examples 164-203.

210. A kit for identifying an individual having cancer who is likely to benefit from treatment with a PD-L1 axis binding antagonist, the kit comprising reagents for determining the expression level of two or more of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein the individual is identified as an individual who is likely to benefit from treatment with a PD-L1 axis binding antagonist if the immune score expression level of the two or more genes is greater than a reference immune score expression level for the two or more genes.

211. A kit for identifying an individual having cancer who is likely to benefit from treatment with a PD-L1 axis binding antagonist, the kit comprising reagents for determining the expression level of one or more of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein the individual is identified as an individual who is likely to benefit from treatment with a PD-L1 axis binding antagonist where the immune score expression level of the one or more genes is greater than a reference immune score expression level for the one or more genes, wherein the benefit comprises an increase in the Overall Survival (OS) of the individual as compared to treatment without the PD-L1 axis binding antagonist.

212. A kit for identifying an individual having cancer who is likely to benefit from treatment with a PD-L1 axis binding antagonist, the kit comprising reagents for determining the expression level of one or more of the genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7 and IGLL5 in a sample from the individual, wherein the individual is identified as an individual who is likely to benefit from treatment with a PD-L1 axis binding antagonist if the immune score expression level of the one or more genes is greater than a reference immune score expression level for the one or more genes, wherein the benefit comprises an extended total survival (OS) of the individual as compared to treatment without the PD-L1 axis binding antagonist.

213. A kit for identifying an individual having cancer who is likely to benefit from treatment with a binding antagonist for the PD-L1 axis, the kit comprising reagents for determining the presence of a Tertiary Lymphoid Structure (TLS) in a tumor sample from the individual, wherein the presence of TLS in the tumor sample identifies the individual as an individual who is likely to benefit from treatment with a binding antagonist for the PD-L1 axis.

214. A kit for identifying an individual having cancer who is likely to benefit from treatment comprising a PD-L1 axis binding antagonist, the kit comprising reagents for determining the expression level of two or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual, wherein the individual is identified as an individual who is likely to benefit from treatment comprising a PD-L1 axis binding antagonist if the immune score expression level of the two or more genes is higher than a reference immune score expression level of the two or more genes.

215. A kit for identifying an individual having cancer who is likely to benefit from treatment with a PD-L1 axis binding antagonist, the kit comprising reagents for determining the number of B cells in a tumor sample from the individual, wherein the individual is identified as an individual who is likely to benefit from treatment with a PD-L1 axis binding antagonist if the number of B cells in the tumor sample is higher than a reference number of B cells, optionally wherein the B cells comprise plasma cells.

216. A kit for identifying an individual having cancer who is likely to benefit from treatment with a PD-L1 axis binding antagonist, the kit comprising reagents for determining whether the individual has clonally expanded B cells in a tumor sample from the individual, wherein having clonally expanded B cells in the sample identifies the individual as an individual who is likely to benefit from treatment with a PD-L1 axis binding antagonist.

217. A PD-L1 axis binding antagonist for use in a method of treating an individual having cancer, the method comprising:

(a) Determining the expression level of two or more of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein the determination of the immune score expression level of the two or more genes in the sample is higher than a reference immune score expression level of the two or more genes; and

218. A PD-L1 axis binding antagonist for use in treating cancer in an individual, wherein the individual has been determined to have an immune score expression level of two or more of the genes CD79A, CD19, BANK1, jcha in, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual that is greater than a reference immune score expression level for the two or more genes.

219. A PD-L1 axis binding antagonist for use in a method of treating an individual having cancer, the method comprising:

220. A PD-L1 axis binding antagonist for use in treating cancer in an individual, wherein the individual has been determined to have an immune score expression level of one or more of the genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual that is greater than a reference immune score expression level for the one or more genes, wherein the individual is identified as an individual likely to benefit from treatment with the PD-L1 axis binding antagonist, wherein the benefit comprises an increased OS in the individual as compared to treatment without the PD-L1 axis binding antagonist, if the immune score expression level of the one or more genes is greater than the reference immune score expression level for the one or more genes.

221. A PD-L1 axis binding antagonist for use in a method of treating an individual having cancer, the method comprising:

222. A PD-L1 axis binding antagonist for use in treating cancer in an individual, wherein the individual has been determined to have an immune score expression level of one or more of the genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from the individual that is higher than a reference immune score expression level of the one or more genes.

223. A PD-L1 axis binding antagonist for use in a method of treating an individual having cancer, the method comprising:

224. A PD-L1 axis binding antagonist for use in treating cancer in an individual, wherein the individual has been determined to have in a sample from the individual an immune score expression level of one or more of the genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 that is higher than a reference immune score expression level for the one or more genes, wherein in the event that the immune score expression level of the one or more genes is higher than the reference immune score expression level for the one or more genes, identifying the individual as an individual likely to comprise treatment with the PD-L1 axis binding antagonist, wherein the benefit comprises an extension of the individual's OS as compared to treatment without the PD-L1 axis binding antagonist.

225. A PD-L1 axis binding antagonist for use in a method of treating an individual having cancer, the method comprising:

(a) Determining the presence of TLS in a tumor sample from the individual; and

226. A PD-L1 axis binding antagonist for use in treating cancer in an individual, wherein the individual has been determined to have TLS present in a tumor sample from the individual.

227. A PD-L1 axis binding antagonist for use in a method of treating an individual having cancer, the method comprising:

228. A PD-L1 axis binding antagonist for use in treating a cancer in an individual, wherein the individual has been determined to have an immune score expression level of two or more of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual that is higher than a reference immune score expression level of the two or more genes.

229. A PD-L1 axis binding antagonist for use in a method of treating an individual having cancer, the method comprising:

(a) Determining the number of B cells in a tumor sample from the individual, optionally wherein the B cells comprise plasma cells; and

230. A PD-L1 axis binding antagonist for use in treating cancer in an individual, wherein the individual has been determined to have a number of B cells in a tumor sample from the individual that is higher than a reference number of B cells, optionally wherein the B cells comprise plasma cells.

231. A PD-L1 axis binding antagonist for use in a method of treating an individual having cancer, the method comprising:

232. A PD-L1 axis binding antagonist for use in treating cancer in an individual, wherein the individual has been determined to have clonally expanded B cells in a tumor sample from the individual.

233. A kit according to any one of embodiments 208 to 216 or a PD-L1 axis binding antagonist for use according to any one of embodiments 217 to 232, wherein the PD-L1 axis binding antagonist is an anti-PD-L1 antagonist antibody, optionally wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain variable region comprising the amino acid sequence of SEQ ID NO 7; and a light chain variable region comprising the amino acid sequence of SEQ ID NO:8, optionally wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO 9; and a light chain comprising the amino acid sequence of SEQ ID NO:10, optionally wherein the anti-PD-L1 antagonist antibody is atezumab.

Sequence listing

<110> Gene Tak Co., ltd (Genentech, inc.)

<120> methods and compositions for cancer immunotherapy

<130> 50474-209WO2

<150> US 63/038,559

<151> 2020-06-12

<160> 83

<170> PatentIn version 3.5

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tcatccagaa tgtgaacaag agccatgggg gcatatacgt gtgccgggtc caggagggca 360

acgagtcata ccagcagtcc tgcggcacct acctccgcgt gcgccagccg ccccccaggc 420

ccttcctgga catgggggag ggcaccaaga accgaatcat cacagccgag gggatcatcc 480

tcctgttctg cgcggtggtg cctgggacgc tgctgctgtt caggaaacga tggcagaacg 540

agaagctcgg gttggatgcc ggggatgaat atgaagatga aaacctttat gaaggcctga 600

acctggacga ctgctccatg tatgaggaca tctcccgggg cctccagggc acctaccagg 660

atgtgggcag cctcaacata ggagatgtcc agctggagaa gccgtgacac ccctactcct 720

gccaggctgc ccccgcctgc tgtgcaccca gctccagtgt ctcagctcac ttccctggga 780

cattctcctt tcagcccttc tgggggcttc cttagtcata ttcccccagt ggggggtggg 840

agggtaacct cactcttctc caggccaggc ctccttggac tcccctgggg gtgtcccact 900

cttcttccct ctaaactgcc ccacctccta acctaatccc cccgccccgc tgcctttccc 960

aggctcccct caccccagcg ggtaatgagc ccttaatcgc tgcctctagg ggagctgatt 1020

gtagcagcct cgttagtgtc accccctcct ccctgatctg tcagggccac ttagtgataa 1080

taaattcttc ccaactgca 1099

<210> 14

<211> 226

<212> PRT

<213> Intelligent people

<400> 14

Met Pro Gly Gly Pro Gly Val Leu Gln Ala Leu Pro Ala Thr Ile Phe

1 5 10 15

Leu Leu Phe Leu Leu Ser Ala Val Tyr Leu Gly Pro Gly Cys Gln Ala

20 25 30

Leu Trp Met His Lys Val Pro Ala Ser Leu Met Val Ser Leu Gly Glu

35 40 45

Asp Ala His Phe Gln Cys Pro His Asn Ser Ser Asn Asn Ala Asn Val

50 55 60

Thr Trp Trp Arg Val Leu His Gly Asn Tyr Thr Trp Pro Pro Glu Phe

65 70 75 80

Leu Gly Pro Gly Glu Asp Pro Asn Gly Thr Leu Ile Ile Gln Asn Val

85 90 95

Asn Lys Ser His Gly Gly Ile Tyr Val Cys Arg Val Gln Glu Gly Asn

100 105 110

Glu Ser Tyr Gln Gln Ser Cys Gly Thr Tyr Leu Arg Val Arg Gln Pro

115 120 125

Pro Pro Arg Pro Phe Leu Asp Met Gly Glu Gly Thr Lys Asn Arg Ile

130 135 140

Ile Thr Ala Glu Gly Ile Ile Leu Leu Phe Cys Ala Val Val Pro Gly

145 150 155 160

Thr Leu Leu Leu Phe Arg Lys Arg Trp Gln Asn Glu Lys Leu Gly Leu

165 170 175

Asp Ala Gly Asp Glu Tyr Glu Asp Glu Asn Leu Tyr Glu Gly Leu Asn

180 185 190

Leu Asp Asp Cys Ser Met Tyr Glu Asp Ile Ser Arg Gly Leu Gln Gly

195 200 205

Thr Tyr Gln Asp Val Gly Ser Leu Asn Ile Gly Asp Val Gln Leu Glu

210 215 220

Lys Pro

225

<210> 15

<211> 2707

<212> DNA

<213> Intelligent people

<400> 15

gtaataccta agagggaagt ggcttcattt cagtggctga cttccagaga gcaatatggc 60

tggttcccca acatgcctca ccctcatcta tatcctttgg cagctcacag ggtcagcagc 120

ctctggaccc gtgaaagagc tggtcggttc cgttggtggg gccgtgactt tccccctgaa 180

gtccaaagta aagcaagttg actctattgt ctggaccttc aacacaaccc ctcttgtcac 240

catacagcca gaagggggca ctatcatagt gacccaaaat cgtaataggg agagagtaga 300

cttcccagat ggaggctact ccctgaagct cagcaaactg aagaagaatg actcagggat 360

ctactatgtg gggatataca gctcatcact ccagcagccc tccacccagg agtacgtgct 420

gcatgtctac gagcacctgt caaagcctaa agtcaccatg ggtctgcaga gcaataagaa 480

tggcacctgt gtgaccaatc tgacatgctg catggaacat ggggaagagg atgtgattta 540

tacctggaag gccctggggc aagcagccaa tgagtcccat aatgggtcca tcctccccat 600

ctcctggaga tggggagaaa gtgatatgac cttcatctgc gttgccagga accctgtcag 660

cagaaacttc tcaagcccca tccttgccag gaagctctgt gaaggtgctg ctgatgaccc 720

agattcctcc atggtcctcc tgtgtctcct gttggtgccc ctcctgctca gtctctttgt 780

actggggcta tttctttggt ttctgaagag agagagacaa gaagagtaca ttgaagagaa 840

gaagagagtg gacatttgtc gggaaactcc taacatatgc ccccattctg gagagaacac 900

agagtacgac acaatccctc acactaatag aacaatccta aaggaagatc cagcaaatac 960

ggtttactcc actgtggaaa taccgaaaaa gatggaaaat ccccactcac tgctcacgat 1020

gccagacaca ccaaggctat ttgcctatga gaatgttatc tagacagcag tgcactcccc 1080

taagtctctg ctcaaaaaaa aaacaattct cggcccaaag aaaacaatca gaagaattca 1140

ctgatttgac tagaaacatc aaggaagaat gaagaacgtt gacttttttc caggataaat 1200

tatctctgat gcttctttag atttaagagt tcataattcc atccactgct gagaaatctc 1260

ctcaaaccca gaaggtttaa tcacttcatc ccaaaaatgg gattgtgaat gtcagcaaac 1320

cataaaaaaa gtgcttagaa gtattcctat aaaaatgtaa atgcaaggtc acacatatta 1380

atgacagcct gttgtattaa tgatggctcc aggtcagtgt ctggagtttc attccatccc 1440

agggcttgga tgtcaggatt ataccaagag tcttgctacc aggagggcaa gaagaccaaa 1500

acagacagac aagtccagca gaagcagatg cacctgacaa aaatggatgt attaattggc 1560

tctataaact atgtgcccag cactatgctg agcttacact aattggtcag acatgctgtc 1620

tgccctcatg aaattggctc caaatgaatg aactactttc atgagcagtt gtagcaggcc 1680

tgaccacaga ttcccagagg gccaggtgtg gatccacagg acttgaaggt caaagttcac 1740

aaagatgaag aatcagggta gctgaccatg tttggcagat actataatgg agacacagaa 1800

gtgtgcatgg cccaaggaca aggacctcca gccaggcttc atttatgcac ttgtgctgca 1860

aaagaaaagt ctaggtttta aggctgtgcc agaacccatc ccaataaaga gaccgagtct 1920

gaagtcacat tgtaaatcta gtgtaggaga cttggagtca ggcagtgaga ctggtggggc 1980

acggggggca gtgggtactt gtaaaccttt aaagatggtt aattcattca atagatattt 2040

attaagaacc tatgcggccc ggcatggtgg ctcacacctg taatcccagc actttgggag 2100

gccaaggtgg gtgggtcatc tgaggtcagg agttcaagac cagcctggcc aacatggtga 2160

aaccccatct ctactaaaga tacaaaaatt tgctgagcgt ggtggtgtgc acctgtaatc 2220

ccagctactc gagaggccaa ggcatgagaa tcgcttgaac ctgggaggtg gaggttgcag 2280

tgagctgaga tggcaccact gcactccggc ctaggcaacg agagcaaaac tccaatacaa 2340

acaaacaaac aaacacctgt gctaggtcag tctggcacgt aagatgaaca tccctaccaa 2400

tacagagctc accatctctt atacttaagt gaaaaacatg gggaagggga aaggggaatg 2460

gctgcttttg atatgttccc tgacacatat cttgaatgga gacctcccta ccaagtgatg 2520

aaagtgttga aaaacttaat aacaaatgct tgttgggcaa gaatgggatt gaggattatc 2580

ttctctcaga aaggcattgt gaaggaattg agccagatct ctctccctac tgcaaaaccc 2640

tattgtagta aaaaagtctt ctttactatc ttaataaaac agatattgtg agattcacat 2700

acaaaaa 2707

<210> 16

<211> 335

<212> PRT

<213> Intelligent people

<400> 16

Met Ala Gly Ser Pro Thr Cys Leu Thr Leu Ile Tyr Ile Leu Trp Gln

1 5 10 15

Leu Thr Gly Ser Ala Ala Ser Gly Pro Val Lys Glu Leu Val Gly Ser

20 25 30

Val Gly Gly Ala Val Thr Phe Pro Leu Lys Ser Lys Val Lys Gln Val

35 40 45

Asp Ser Ile Val Trp Thr Phe Asn Thr Thr Pro Leu Val Thr Ile Gln

50 55 60

Pro Glu Gly Gly Thr Ile Ile Val Thr Gln Asn Arg Asn Arg Glu Arg

65 70 75 80

Val Asp Phe Pro Asp Gly Gly Tyr Ser Leu Lys Leu Ser Lys Leu Lys

85 90 95

Lys Asn Asp Ser Gly Ile Tyr Tyr Val Gly Ile Tyr Ser Ser Ser Leu

100 105 110

Gln Gln Pro Ser Thr Gln Glu Tyr Val Leu His Val Tyr Glu His Leu

115 120 125

Ser Lys Pro Lys Val Thr Met Gly Leu Gln Ser Asn Lys Asn Gly Thr

130 135 140

Cys Val Thr Asn Leu Thr Cys Cys Met Glu His Gly Glu Glu Asp Val

145 150 155 160

Ile Tyr Thr Trp Lys Ala Leu Gly Gln Ala Ala Asn Glu Ser His Asn

165 170 175

Gly Ser Ile Leu Pro Ile Ser Trp Arg Trp Gly Glu Ser Asp Met Thr

180 185 190

Phe Ile Cys Val Ala Arg Asn Pro Val Ser Arg Asn Phe Ser Ser Pro

195 200 205

Ile Leu Ala Arg Lys Leu Cys Glu Gly Ala Ala Asp Asp Pro Asp Ser

210 215 220

Ser Met Val Leu Leu Cys Leu Leu Leu Val Pro Leu Leu Leu Ser Leu

225 230 235 240

Phe Val Leu Gly Leu Phe Leu Trp Phe Leu Lys Arg Glu Arg Gln Glu

245 250 255

Glu Tyr Ile Glu Glu Lys Lys Arg Val Asp Ile Cys Arg Glu Thr Pro

260 265 270

Asn Ile Cys Pro His Ser Gly Glu Asn Thr Glu Tyr Asp Thr Ile Pro

275 280 285

His Thr Asn Arg Thr Ile Leu Lys Glu Asp Pro Ala Asn Thr Val Tyr

290 295 300

Ser Thr Val Glu Ile Pro Lys Lys Met Glu Asn Pro His Ser Leu Leu

305 310 315 320

Thr Met Pro Asp Thr Pro Arg Leu Phe Ala Tyr Glu Asn Val Ile

325 330 335

<210> 17

<211> 2611

<212> DNA

<213> Intelligent

<400> 17

aactgagtgg ctgtgaaagg gtggggtttg ctcagactgt ccttcctctc tggactgtaa 60

gaatatgtct ccagggccag tgtctgctgc gatcgagtcc caccttccaa gtcctggcat 120

ctcaatgcat ctgggaagct acctgcatta agtcaggact gagcacacag gtgaactcca 180

gaaagaagaa gctatggccg cagtgattct ggagagcatc tttctgaagc gatcccaaca 240

gaaaaagaaa acatcacctc taaacttcaa gaagcgcctg tttctcttga ccgtgcacaa 300

actctcctac tatgagtatg actttgaacg tgggagaaga ggcagtaaga agggttcaat 360

agatgttgag aagatcactt gtgttgaaac agtggttcct gaaaaaaatc ctcctccaga 420

aagacagatt ccgagaagag gtgaagagtc cagtgaaatg gagcaaattt caatcattga 480

aaggttccct tatcccttcc aggttgtata tgatgaaggg cctctctacg tcttctcccc 540

aactgaagaa ctaaggaagc ggtggattca ccagctcaaa aacgtaatcc ggtacaacag 600

tgatctggtt cagaaatatc acccttgctt ctggatcgat gggcagtatc tctgctgctc 660

tcagacagcc aaaaatgcta tgggctgcca aattttggag aacaggaatg gaagcttaaa 720

acctgggagt tctcaccgga agacaaaaaa gcctcttccc ccaacgcctg aggaggacca 780

gatcttgaaa aagccactac cgcctgagcc agcagcagca ccagtctcca caagtgagct 840

gaaaaaggtt gtggcccttt atgattacat gccaatgaat gcaaatgatc tacagctgcg 900

gaagggtgat gaatatttta tcttggagga aagcaactta ccatggtgga gagcacgaga 960

taaaaatggg caggaaggct acattcctag taactatgtc actgaagcag aagactccat 1020

agaaatgtat gagtggtatt ccaaacacat gactcggagt caggctgagc aactgctaaa 1080

gcaagagggg aaagaaggag gtttcattgt cagagactcc agcaaagctg gcaaatatac 1140

agtgtctgtg tttgctaaat ccacagggga ccctcaaggg gtgatacgtc attatgttgt 1200

gtgttccaca cctcagagcc agtattacct ggctgagaag caccttttca gcaccatccc 1260

tgagctcatt aactaccatc agcacaactc tgcaggactc atatccaggc tcaaatatcc 1320

agtgtctcaa caaaacaaga atgcaccttc cactgcaggc ctgggatacg gatcatggga 1380

aattgatcca aaggacctga ccttcttgaa ggagctgggg actggacaat ttggggtagt 1440

gaagtatggg aaatggagag gccagtacga cgtggccatc aagatgatca aagaaggctc 1500

catgtctgaa gatgaattca ttgaagaagc caaagtcatg atgaatcttt cccatgagaa 1560

gctggtgcag ttgtatggcg tctgcaccaa gcagcgcccc atcttcatca tcactgagta 1620

catggccaat ggctgcctcc tgaactacct gagggagatg cgccaccgct tccagactca 1680

gcagctgcta gagatgtgca aggatgtctg tgaagccatg gaatacctgg agtcaaagca 1740

gttccttcac cgagacctgg cagctcgaaa ctgtttggta aacgatcaag gagttgttaa 1800

agtatctgat ttcggcctgt ccaggtatgt cctggatgat gaatacacaa gctcagtagg 1860

ctccaaattt ccagtccggt ggtccccacc ggaagtcctg atgtatagca agttcagcag 1920

caaatctgac atttgggctt ttggggtttt gatgtgggaa atttactccc tggggaagat 1980

gccatatgag agatttacta acagtgagac tgctgaacac attgcccaag gcctacgtct 2040

ctacaggcct catctggctt cagagaaggt atataccatc atgtacagtt gctggcatga 2100

gaaagcagat gagcgtccca ctttcaaaat tcttctgagc aatattctag atgtcatgga 2160

tgaagaatcc tgagctcgcc aataagcttc ttggttctac ttctcttctc cacaagcccc 2220

aatttcactt tctcagagga aatcccaagc ttaggagccc tggagccttt gtgctcccac 2280

tcaatacaaa aaggcccctc tctacatctg ggaatgcacc tcttctttga ttccctggga 2340

tagtggcttc tgagcaaagg ccaagaaatt attgtgcctg aaatttcccg agagaattaa 2400

gacagactga atttgcgatg aaaatatttt ttaggaggga ggatgtaaat agccgcacaa 2460

aggggtccaa cagctctttg agtaggcatt tggtagagct tgggggtgtg tgtgtggggg 2520

tggaccgaat ttggcaagaa tgaaatggtg tcataaagat gggaggggag ggtgttttga 2580

taaaataaaa ttactagaaa gcttgaaagt c 2611

<210> 18

<211> 659

<212> PRT

<213> Intelligent

<400> 18

Met Ala Ala Val Ile Leu Glu Ser Ile Phe Leu Lys Arg Ser Gln Gln

1 5 10 15

Lys Lys Lys Thr Ser Pro Leu Asn Phe Lys Lys Arg Leu Phe Leu Leu

20 25 30

Thr Val His Lys Leu Ser Tyr Tyr Glu Tyr Asp Phe Glu Arg Gly Arg

35 40 45

Arg Gly Ser Lys Lys Gly Ser Ile Asp Val Glu Lys Ile Thr Cys Val

50 55 60

Glu Thr Val Val Pro Glu Lys Asn Pro Pro Pro Glu Arg Gln Ile Pro

65 70 75 80

Arg Arg Gly Glu Glu Ser Ser Glu Met Glu Gln Ile Ser Ile Ile Glu

85 90 95

Arg Phe Pro Tyr Pro Phe Gln Val Val Tyr Asp Glu Gly Pro Leu Tyr

100 105 110

Val Phe Ser Pro Thr Glu Glu Leu Arg Lys Arg Trp Ile His Gln Leu

115 120 125

Lys Asn Val Ile Arg Tyr Asn Ser Asp Leu Val Gln Lys Tyr His Pro

130 135 140

Cys Phe Trp Ile Asp Gly Gln Tyr Leu Cys Cys Ser Gln Thr Ala Lys

145 150 155 160

Asn Ala Met Gly Cys Gln Ile Leu Glu Asn Arg Asn Gly Ser Leu Lys

165 170 175

Pro Gly Ser Ser His Arg Lys Thr Lys Lys Pro Leu Pro Pro Thr Pro

180 185 190

Glu Glu Asp Gln Ile Leu Lys Lys Pro Leu Pro Pro Glu Pro Ala Ala

195 200 205

Ala Pro Val Ser Thr Ser Glu Leu Lys Lys Val Val Ala Leu Tyr Asp

210 215 220

Tyr Met Pro Met Asn Ala Asn Asp Leu Gln Leu Arg Lys Gly Asp Glu

225 230 235 240

Tyr Phe Ile Leu Glu Glu Ser Asn Leu Pro Trp Trp Arg Ala Arg Asp

245 250 255

Lys Asn Gly Gln Glu Gly Tyr Ile Pro Ser Asn Tyr Val Thr Glu Ala

260 265 270

Glu Asp Ser Ile Glu Met Tyr Glu Trp Tyr Ser Lys His Met Thr Arg

275 280 285

Ser Gln Ala Glu Gln Leu Leu Lys Gln Glu Gly Lys Glu Gly Gly Phe

290 295 300

Ile Val Arg Asp Ser Ser Lys Ala Gly Lys Tyr Thr Val Ser Val Phe

305 310 315 320

Ala Lys Ser Thr Gly Asp Pro Gln Gly Val Ile Arg His Tyr Val Val

325 330 335

Cys Ser Thr Pro Gln Ser Gln Tyr Tyr Leu Ala Glu Lys His Leu Phe

340 345 350

Ser Thr Ile Pro Glu Leu Ile Asn Tyr His Gln His Asn Ser Ala Gly

355 360 365

Leu Ile Ser Arg Leu Lys Tyr Pro Val Ser Gln Gln Asn Lys Asn Ala

370 375 380

Pro Ser Thr Ala Gly Leu Gly Tyr Gly Ser Trp Glu Ile Asp Pro Lys

385 390 395 400

Asp Leu Thr Phe Leu Lys Glu Leu Gly Thr Gly Gln Phe Gly Val Val

405 410 415

Lys Tyr Gly Lys Trp Arg Gly Gln Tyr Asp Val Ala Ile Lys Met Ile

420 425 430

Lys Glu Gly Ser Met Ser Glu Asp Glu Phe Ile Glu Glu Ala Lys Val

435 440 445

Met Met Asn Leu Ser His Glu Lys Leu Val Gln Leu Tyr Gly Val Cys

450 455 460

Thr Lys Gln Arg Pro Ile Phe Ile Ile Thr Glu Tyr Met Ala Asn Gly

465 470 475 480

Cys Leu Leu Asn Tyr Leu Arg Glu Met Arg His Arg Phe Gln Thr Gln

485 490 495

Gln Leu Leu Glu Met Cys Lys Asp Val Cys Glu Ala Met Glu Tyr Leu

500 505 510

Glu Ser Lys Gln Phe Leu His Arg Asp Leu Ala Ala Arg Asn Cys Leu

515 520 525

Val Asn Asp Gln Gly Val Val Lys Val Ser Asp Phe Gly Leu Ser Arg

530 535 540

Tyr Val Leu Asp Asp Glu Tyr Thr Ser Ser Val Gly Ser Lys Phe Pro

545 550 555 560

Val Arg Trp Ser Pro Pro Glu Val Leu Met Tyr Ser Lys Phe Ser Ser

565 570 575

Lys Ser Asp Ile Trp Ala Phe Gly Val Leu Met Trp Glu Ile Tyr Ser

580 585 590

Leu Gly Lys Met Pro Tyr Glu Arg Phe Thr Asn Ser Glu Thr Ala Glu

595 600 605

His Ile Ala Gln Gly Leu Arg Leu Tyr Arg Pro His Leu Ala Ser Glu

610 615 620

Lys Val Tyr Thr Ile Met Tyr Ser Cys Trp His Glu Lys Ala Asp Glu

625 630 635 640

Arg Pro Thr Phe Lys Ile Leu Leu Ser Asn Ile Leu Asp Val Met Asp

645 650 655

Glu Glu Ser

<210> 19

<211> 891

<212> DNA

<213> Intelligent people

<400> 19

attgttctca acattctagc tgctcttgct gcatttgctc tggaattctt gtagagatat 60

tacttgtcct tccaggctgt tctttctgta gctcccttgt tttctttttg tgatcatgtt 120

gcagatggct gggcagtgct cccaaaatga atattttgac agtttgttgc atgcttgcat 180

accttgtcaa cttcgatgtt cttctaatac tcctcctcta acatgtcagc gttattgtaa 240

tgcaagtgtg accaattcag tgaaaggaac gaatgcgatt ctctggacct gtttgggact 300

gagcttaata atttctttgg cagttttcgt gctaatgttt ttgctaagga agataaactc 360

tgaaccatta aaggacgagt ttaaaaacac aggatcaggt ctcctgggca tggctaacat 420

tgacctggaa aagagcagga ctggtgatga aattattctt ccgagaggcc tcgagtacac 480

ggtggaagaa tgcacctgtg aagactgcat caagagcaaa ccgaaggtcg actctgacca 540

ttgctttcca ctcccagcta tggaggaagg cgcaaccatt cttgtcacca cgaaaacgaa 600

tgactattgc aagagcctgc cagctgcttt gagtgctacg gagatagaga aatcaatttc 660

tgctaggtaa ttaaccattt cgactcgagc agtgccactt taaaaatctt ttgtcagaat 720

agatgatgtg tcagatctct ttaggatgac tgtatttttc agttgccgat acagcttttt 780

gtcctctaac tgtggaaact ctttatgtta gatatatttc tctaggttac tgttgggagc 840

ttaatggtag aaacttcctt ggtttcatga ttaaactctt ttttttcctg a 891

<210> 20

<211> 184

<212> PRT

<213> Intelligent people

<400> 20

Met Leu Gln Met Ala Gly Gln Cys Ser Gln Asn Glu Tyr Phe Asp Ser

1 5 10 15

Leu Leu His Ala Cys Ile Pro Cys Gln Leu Arg Cys Ser Ser Asn Thr

20 25 30

Pro Pro Leu Thr Cys Gln Arg Tyr Cys Asn Ala Ser Val Thr Asn Ser

35 40 45

Val Lys Gly Thr Asn Ala Ile Leu Trp Thr Cys Leu Gly Leu Ser Leu

50 55 60

Ile Ile Ser Leu Ala Val Phe Val Leu Met Phe Leu Leu Arg Lys Ile

65 70 75 80

Asn Ser Glu Pro Leu Lys Asp Glu Phe Lys Asn Thr Gly Ser Gly Leu

85 90 95

Leu Gly Met Ala Asn Ile Asp Leu Glu Lys Ser Arg Thr Gly Asp Glu

100 105 110

Ile Ile Leu Pro Arg Gly Leu Glu Tyr Thr Val Glu Glu Cys Thr Cys

115 120 125

Glu Asp Cys Ile Lys Ser Lys Pro Lys Val Asp Ser Asp His Cys Phe

130 135 140

Pro Leu Pro Ala Met Glu Glu Gly Ala Thr Ile Leu Val Thr Thr Lys

145 150 155 160

Thr Asn Asp Tyr Cys Lys Ser Leu Pro Ala Ala Leu Ser Ala Thr Glu

165 170 175

Ile Glu Lys Ser Ile Ser Ala Arg

180

<210> 21

<211> 1286

<212> DNA

<213> Intelligent people

<400> 21

agaagaagtg aagtcaagat gaagaaccat ttgcttttct ggggagtcct ggcggttttt 60

attaaggctg ttcatgtgaa agcccaagaa gatgaaagga ttgttcttgt tgacaacaaa 120

tgtaagtgtg cccggattac ttccaggatc atccgttctt ccgaagatcc taatgaggac 180

attgtggaga gaaacatccg aattattgtt cctctgaaca acagggagaa tatctctgat 240

cccacctcac cattgagaac cagatttgtg taccatttgt ctgacctctg taaaaaatgt 300

gatcctacag aagtggagct ggataatcag atagttactg ctacccagag caatatctgt 360

gatgaagaca gtgctacaga gacctgctac acttatgaca gaaacaagtg ctacacagct 420

gtggtcccac tcgtatatgg tggtgagacc aaaatggtgg aaacagcctt aaccccagat 480

gcctgctatc ctgactaatt taagtcattg ctgactgcat agctcttttt cttgagaggc 540

tctccatttt gattcagaaa gttagcatat ttattaccaa tgaatttgaa accagggctt 600

tttttttttt ttgggtgatg taaaaccaac tccctgccac caaaataatt aaaatagtca 660

cattgttatc tttattaggt aatcacttct taattatatg ttcatactct aagtatcaaa 720

atcttccaat tatcatgctc acctgaaaga ggtatgctct cttaggaata cagtttctag 780

cattaaacaa ataaacaagg ggagaaaata aaactcaagg actgaaaatc aggaggtgta 840

ataaaatgtt cctcgcattc ccccccgctt tttttttttt ttttgacttt gccttggaga 900

gccagagctt ccgcattttc tttactattc tttttaaaaa aagtttcact gtgtagagaa 960

catatatgca taaacatagg tcaattatat gtctccatta gaaaaataat aattggaaaa 1020

catgttctag aactagttac aaaaataatt taaggtgaaa tctctaatat ttataaaagt 1080

agcaaaataa atgcataatt aaaatatatt tggacataac agacttggaa gcagatgata 1140

cagacttctt tttttcataa tcaggttagt gtaagaaatt gccatttgaa acaatccatt 1200

ttgtaactga accttatgaa atatatgtat ttcatggtac gtattctcta gcacagtctg 1260

agcaattaaa tagattcata agcata 1286

<210> 22

<211> 159

<212> PRT

<213> Intelligent

<400> 22

Met Lys Asn His Leu Leu Phe Trp Gly Val Leu Ala Val Phe Ile Lys

1 5 10 15

Ala Val His Val Lys Ala Gln Glu Asp Glu Arg Ile Val Leu Val Asp

20 25 30

Asn Lys Cys Lys Cys Ala Arg Ile Thr Ser Arg Ile Ile Arg Ser Ser

35 40 45

Glu Asp Pro Asn Glu Asp Ile Val Glu Arg Asn Ile Arg Ile Ile Val

50 55 60

Pro Leu Asn Asn Arg Glu Asn Ile Ser Asp Pro Thr Ser Pro Leu Arg

65 70 75 80

Thr Arg Phe Val Tyr His Leu Ser Asp Leu Cys Lys Lys Cys Asp Pro

85 90 95

Thr Glu Val Glu Leu Asp Asn Gln Ile Val Thr Ala Thr Gln Ser Asn

100 105 110

Ile Cys Asp Glu Asp Ser Ala Thr Glu Thr Cys Tyr Thr Tyr Asp Arg

115 120 125

Asn Lys Cys Tyr Thr Ala Val Val Pro Leu Val Tyr Gly Gly Glu Thr

130 135 140

Lys Met Val Glu Thr Ala Leu Thr Pro Asp Ala Cys Tyr Pro Asp

145 150 155

<210> 23

<211> 1300

<212> DNA

<213> Intelligent

<400> 23

gtgaccgctt cagggcagtt ggtagatgcc cctctgggag agatccccag gggtgacagc 60

catggaccct ggaagggcct gggctaggga cagggaccag agccagtcca gggagaggac 120

agagccaatg gactggggtg tactgtaaca gccctgctgg cgagagggac cagggcaccg 180

tcctccaggg agcccatgct gcaagtcggg ccagaggtgc ccctgaacct gaaggccaat 240

gagacccaag acaggccaag tgggttgtga gacccctgag gagctgggcc ctggtcccag 300

gcagcgctgg cccctgctgc tgctgggtct ggccatggtc gcccatggcc tgctgcgccc 360

aatggttgca ccgcaaagcg gggacccaga ccctggagcc tcagttggaa gcagccgatc 420

cagcctgcgg agcctgtggg gcaggctcct gctccagccc agcccccaga gagcagaccc 480

caggtgctgg ccccgggggt tttggtctga gcctcagtca ctgtgttatg tcttcggaac 540

tgggaccaag gtcaccgtcc taggtcagcc caaggccaac cccactgtca ctctgttccc 600

gccctcctct gaggagctcc aagccaacaa ggccacacta gtgtgtctga tcagtgactt 660

ctacccggga gctgtgacag tggcctggaa ggcagatggc agccccgtca aggcgggagt 720

ggagaccacc aaaccctcca aacagagcaa caacaagtac gcggccagca gctacctgag 780

cctgacgccc gagcagtgga agtcccacag aagctacagc tgccaggtca cgcatgaagg 840

gagcaccgtg gagaagacag tggcccctac agaatgttca taggttccca actctaaccc 900

cacccacggg agcctggagc tgcaggatcc caggggaggg gtctctctcc ccatcccaag 960

tcatccagcc cttctccctg cactcatgaa accccaataa atatcctcat tgacaaccag 1020

aaatcttgtt ttatctcatt ttttttctca cataaattgc tagcctcccc ggggttctca 1080

gtgtggggta cagggaattc tgcacccagt gtgaaaatca cccaagggag gaggctcaca 1140

gcctccctga gtcatctccc cagagggtcc ttcctctccc agtcacccct tctccaactc 1200

tccactgtac ccctgagcta ccagtctggc atcagttcag accagtccca caccctccta 1260

aattttactt ctcaataaat acctgatcat gtaaaacgca 1300

<210> 24

<211> 214

<212> PRT

<213> Intelligent people

<400> 24

Met Arg Pro Lys Thr Gly Gln Val Gly Cys Glu Thr Pro Glu Glu Leu

1 5 10 15

Gly Pro Gly Pro Arg Gln Arg Trp Pro Leu Leu Leu Leu Gly Leu Ala

20 25 30

Met Val Ala His Gly Leu Leu Arg Pro Met Val Ala Pro Gln Ser Gly

35 40 45

Asp Pro Asp Pro Gly Ala Ser Val Gly Ser Ser Arg Ser Ser Leu Arg

50 55 60

Ser Leu Trp Gly Arg Leu Leu Leu Gln Pro Ser Pro Gln Arg Ala Asp

65 70 75 80

Pro Arg Cys Trp Pro Arg Gly Phe Trp Ser Glu Pro Gln Ser Leu Cys

85 90 95

Tyr Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu Gly Gln Pro Lys

100 105 110

Ala Asn Pro Thr Val Thr Leu Phe Pro Pro Ser Ser Glu Glu Leu Gln

115 120 125

Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr Pro Gly

130 135 140

Ala Val Thr Val Ala Trp Lys Ala Asp Gly Ser Pro Val Lys Ala Gly

145 150 155 160

Val Glu Thr Thr Lys Pro Ser Lys Gln Ser Asn Asn Lys Tyr Ala Ala

165 170 175

Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His Arg Ser

180 185 190

Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys Thr Val

195 200 205

Ala Pro Thr Glu Cys Ser

210

<210> 25

<211> 5878

<212> DNA

<213> Intelligent people

<400> 25

attgggtaca tgcgcagtgc cgctcgcgcg ggccgcgcca atcctgcagc gccttcaaca 60

ggtttcgggc ggcgaattcc agttctccgg gttttggggc tccctgcggg aggcagtgct 120

ggatctggga atctctagga aaggaaagag caaaggaggg gaaagatggg gggctgcagt 180

ctccacgtac gtccctcaaa gcgcgtccta aaacccggat aaccggagcg ctccccatgg 240

accacacgga gggctcgccc gcggaggagc cgcctgcgca tgctccatcg cctgggaaat 300

ttggtgagcg gcctccacct aaacgactta ctagggaagc tatgcgaaat tatttaaaag 360

agcgagggga tcaaacagta cttattcttc atgcaaaagt tgcacagaag tcatatggaa 420

atgaaaaaag gtttttttgc ccacctcctt gtgtatatct tatgggcagt ggatggaaga 480

aaaaaaaaga acaaatggaa cgcgatggtt gttctgaaca agagtctcaa ccgtgtgcat 540

ttattgggat aggaaatagt gaccaagaaa tgcagcagct aaacttggaa ggaaagaact 600

attgcacagc caaaacattg tatatatctg actcagacaa gcgaaagcac ttcatgttgt 660

ctgtaaagat gttctatggc aacagtgatg acattggtgt gttcctcagc aagcggataa 720

aagtcatctc caaaccttcc aaaaagaagc agtcattgaa aaatgctgac ttatgcattg 780

cctcaggaac aaaggtggct ctgtttaatc gactacgatc ccagacagtt agtaccagat 840

acttgcatgt agaaggaggt aattttcatg ccagttcaca gcagtgggga gcctttttta 900

ttcatctctt ggatgatgat gaatcagaag gagaagaatt cacagtccga gatggctaca 960

tccattatgg acaaacagtc aaacttgtgt gctcagttac tggcatggca ctcccaagat 1020

tgataattag gaaagttgat aagcagaccg cattattgga tgcagatgat cctgtgtcac 1080

aactccataa atgtgcattt taccttaagg atacagaaag aatgtatttg tgcctttctc 1140

aagaaagaat aattcaattt caggccactc catgtccaaa agaaccaaat aaagagatga 1200

taaatgatgg cgcttcctgg acaatcatta gcacagataa ggcagagtat acattttatg 1260

agggaatggg ccctgtcctt gccccagtca ctcctgtgcc tgtggtagag agccttcagt 1320

tgaatggcgg tggggacgta gcaatgcttg aacttacagg acagaatttc actccaaatt 1380

tacgagtgtg gtttggggat gtagaagctg aaactatgta caggtgtgga gagagtatgc 1440

tctgtgtcgt cccagacatt tctgcattcc gagaaggttg gagatgggtc cggcaaccag 1500

tccaggttcc agtaactttg gtccgaaatg atggaatcat ttattccacc agccttacct 1560

ttacctacac accagaacca gggccgcggc cacattgcag tgcagcagga gcaatccttc 1620

gagccaattc aagccaggtg ccccctaacg aatcaaacac aaacagcgag ggaagttaca 1680

caaacgccag cacaaattca accagtgtca catcatctac agccacagtg gtatcctaac 1740

taccgtcttt ttgctaggac ttaaactgac ttgagtgtgg caaaaagtta acaaaaaagg 1800

agaaaaaatg aacaatcgtt tgtggtttct tgggaaaact tttcatacca ggtgatacta 1860

ttcaaaaacc ccgttgtctc cctgcaagtg ctgatttgaa atgcagaagc cacagtaaaa 1920

aaaaaaaaaa aaaaaaaaaa aaagaaaaaa aaatcaaaat gtataaatat tggaaatcaa 1980

gtttttcagc tgttttgttg gttggttggt tggtttttgt ttggttttgt ttaaatgggc 2040

aagaagtaaa taatgtggct ggaatacaag ttgaacaaac tagaagacac aaatctaaca 2100

tagtttttat ggaccaagga acttgtatat tgtataagct ttagtaaaag gtacattttc 2160

accatacctt tttttatatc acggtattat agtacacctt gttaccaaat aggttgttct 2220

cttccccacc cacctttgag cttttgctct aaaatacatt caggttccaa gcctgaccat 2280

ccttgtttaa tctatcatac tcttccaggt tttttttttt tggtctaagg ctggaacttt 2340

tttctttttt ttcagctgaa gtcttatgac ttttcatgag tcaaaattgt ttggatttca 2400

gcaagtcaaa tcttgcaaag gcctgcatat tttttttaag attatatgaa gtctgtgcaa 2460

aagctttaaa aaaatgcctc tgccttgcct gcaatacatg caatgtatgt taacttagtc 2520

tctcttctca gacactgttg gtagttattt ctgtgttttc ctttttttaa aaaaaaatat 2580

ggacttattg tggttatctg agaggttcta acattcacat gcaatttggt gtggccattt 2640

agctattaat gagttaatgg cgcagaactt gttgatattt gaagtgttct ctcccctttt 2700

cccatgacgt aaatacatag gtgtgttcca ggatttgttc aggtttttcc cccctcctaa 2760

tcttgtacat aacttgtatt atgtgtaagt taaacatttt attttgaact tggaatgttc 2820

ccagtgattt cattcagcag ggtattttct gccttgttgg caagtgacaa aaaatatggg 2880

aagtatttgc taccagttgg tagatggtgc ccttaatggt agaatgagga aaatgtccgc 2940

aaaagcatgt tttattatct ttactttttt ggggggttgg agggggtagc ctagccagaa 3000

catcattgta atcttaaaac ataagatgct tttattagat gatcaactaa aatagctgga 3060

agacagtact ttagaaacag atagttgtaa gattataaaa tgcaaatgta acttatgttt 3120

tcattttttt ctctgccttt tttgtttgtt tgttttctct tttccagtac tgagcatctc 3180

cacaaatgtc tcctaactca gaaaatgttt cttttctttt cagttgagat ttggttgcat 3240

tcagggttgt aggttggcct tgcttgctaa ccccgccggt tttaccgtgc tttcattcct 3300

gaactttgtt tatgcctttg tttggtttct tcgaaattgc agcagactca ttgggctaca 3360

tttagtacag gaaccacgtg tgtaatgtta tacaacacag tctagtaata caatcatccc 3420

tcttagagta aaaactacct ctagattgtg gtaagctttt actgtcccat aaaacaggag 3480

ccacagtacc ttatgaatgc aaaactgtaa cttcctacag tgtttcccta cagaacattg 3540

tctttctggt gtcctgggct gttttgaaaa agtttccatt aatagacttt ttagaaatta 3600

ttattagtag catttttttt ccagctttgc tgtcttcatc actcactcta tgctcagact 3660

atgccactgt aaatattctt cctaacatct ttaaatcgcc ttttcctcag ttttcaaggg 3720

gaaggtcatt tgtaaagcac gttaggtggt taaatcagtt attgcggttt tctcttactg 3780

caagcctttt taatcacccc caggctgcat tttattctat atcgcctttt ttcttcaaat 3840

ctgctccaat cactcacttc tctcttataa gctaatcctg cctcacacct taaatctgtt 3900

tcagtgatca agggcagaac tcattgtggc cttatctttc tttgttgtaa ttgttcactg 3960

tctctttctt acagaccact tatttctgag tagtagttat tcctctctat ggagtcatgg 4020

caggaatcat tacacagtgc ttttgttcag agcatggaca tgttcctagt gctgctttgc 4080

tttaacggcc acaagtttcc tccacttcct aggtttggta tttagttaag gaatcatatt 4140

aaattaacca ataacaaaag agatactttt gaagaacaaa ctattcctta cccatttttg 4200

tagctcaaaa ataatttttc aagttcatga ccttattaaa atgaacttgt gtttttttaa 4260

caaacatgta tgttttattt tgatagtttc tttccgtaag ataattgaaa tattatactg 4320

taaacccttt tcttttcttt ttttgaaaag tccaagaatg tacttataca ggcatttttc 4380

cccacctatt tttggccatt ctcataccac agactaaaga gtgaaatgat ttgtccattg 4440

tagcttattg tttatcagta gttcttttgt cagctgctta cattttttct ttcatggttt 4500

tgtgaatcat tttcagtatg taatttatag gaaccttgtc ctctggttat agtagactgt 4560

gtgccctcct ccagtgatgg cattattaga catgctggtc atttaccctc agaaagactc 4620

tcttattaga atggtgagtg cttcagttat agtatgtttg aatttttaaa aaattctgtt 4680

ttagaaatgt atcttatgct ctcatgacta tgcagtttct aaacatacac atagaagctg 4740

agtctctgat ccaatatgtt tttatttgtt ccatttaatt tatcacatag attgggaagg 4800

caagctaaaa gccttaaaaa tgccctttat attttgagtg atttcagcgt tgaacactag 4860

tatactatct aaatttgctg ctcactttct ttaaactgtg gcaattaaag gcatgtttat 4920

acatgactta atcgtgaaat gtttgtcact cttactgcac agacttatct gcaatcataa 4980

ctggttagtt tttttgtttt gttttgtttt attgttttta atgaaactgg taccatctgt 5040

gctttcacaa aaaacttcca atgccatttt tgagaactaa cctaactagt catgctaacc 5100

agaaaatcca ctggggagga ggttcctttt gaaacaaaat gctgttcagt tagtaaccaa 5160

gttactttga ttgcaaaagc agctgtgttt ctgataagta ctgaacaaat gtgtgtaatt 5220

ttctgtgcca gacttatgac tttgttttca agcactgtaa tgtgggatgg atggttagaa 5280

acaataatat attagggttt ctgtttaacc ctttcaggac tgaactgtat ctccttttgt 5340

taattttccc ctgtgttgtg ataaatgttt gccagcattc agtactgtgt tggtccagat 5400

gtaggtttat atgctcattt ttagcttatt tcttgtacct tgcagcatgc tctacgcatt 5460

cagtccttaa ggggtttatt ttacaaactg tgcgcctgta aggtttatta gcaataagat 5520

agaaaattga gcaagtttat accataattt tgtagaaaaa aagaatctgc tcagttccat 5580

atttcatccg tgaaaaactt gcaatacgag cagtttcaag gaataaataa aaaggaaatg 5640

taaaccattg taaaagtctt ctgtcgaatg tgcctgatgc atgtattatc gtcttttatt 5700

tcagaatact tcataaagat aaaattaaat tctatattat agttggtgta tttacaatct 5760

taccatgtac atcacatcaa agtgatagct ctactaattt aatttccttg tcaatgtttt 5820

taactatata gtgctttaaa gagatttttt ttccctgtgt aaaaaaaaaa aaaaaaaa 5878

<210> 26

<211> 500

<212> PRT

<213> Intelligent people

<400> 26

Met Asp His Thr Glu Gly Ser Pro Ala Glu Glu Pro Pro Ala His Ala

1 5 10 15

Pro Ser Pro Gly Lys Phe Gly Glu Arg Pro Pro Pro Lys Arg Leu Thr

20 25 30

Arg Glu Ala Met Arg Asn Tyr Leu Lys Glu Arg Gly Asp Gln Thr Val

35 40 45

Leu Ile Leu His Ala Lys Val Ala Gln Lys Ser Tyr Gly Asn Glu Lys

50 55 60

Arg Phe Phe Cys Pro Pro Pro Cys Val Tyr Leu Met Gly Ser Gly Trp

65 70 75 80

Lys Lys Lys Lys Glu Gln Met Glu Arg Asp Gly Cys Ser Glu Gln Glu

85 90 95

Ser Gln Pro Cys Ala Phe Ile Gly Ile Gly Asn Ser Asp Gln Glu Met

100 105 110

Gln Gln Leu Asn Leu Glu Gly Lys Asn Tyr Cys Thr Ala Lys Thr Leu

115 120 125

Tyr Ile Ser Asp Ser Asp Lys Arg Lys His Phe Met Leu Ser Val Lys

130 135 140

Met Phe Tyr Gly Asn Ser Asp Asp Ile Gly Val Phe Leu Ser Lys Arg

145 150 155 160

Ile Lys Val Ile Ser Lys Pro Ser Lys Lys Lys Gln Ser Leu Lys Asn

165 170 175

Ala Asp Leu Cys Ile Ala Ser Gly Thr Lys Val Ala Leu Phe Asn Arg

180 185 190

Leu Arg Ser Gln Thr Val Ser Thr Arg Tyr Leu His Val Glu Gly Gly

195 200 205

Asn Phe His Ala Ser Ser Gln Gln Trp Gly Ala Phe Phe Ile His Leu

210 215 220

Leu Asp Asp Asp Glu Ser Glu Gly Glu Glu Phe Thr Val Arg Asp Gly

225 230 235 240

Tyr Ile His Tyr Gly Gln Thr Val Lys Leu Val Cys Ser Val Thr Gly

245 250 255

Met Ala Leu Pro Arg Leu Ile Ile Arg Lys Val Asp Lys Gln Thr Ala

260 265 270

Leu Leu Asp Ala Asp Asp Pro Val Ser Gln Leu His Lys Cys Ala Phe

275 280 285

Tyr Leu Lys Asp Thr Glu Arg Met Tyr Leu Cys Leu Ser Gln Glu Arg

290 295 300

Ile Ile Gln Phe Gln Ala Thr Pro Cys Pro Lys Glu Pro Asn Lys Glu

305 310 315 320

Met Ile Asn Asp Gly Ala Ser Trp Thr Ile Ile Ser Thr Asp Lys Ala

325 330 335

Glu Tyr Thr Phe Tyr Glu Gly Met Gly Pro Val Leu Ala Pro Val Thr

340 345 350

Pro Val Pro Val Val Glu Ser Leu Gln Leu Asn Gly Gly Gly Asp Val

355 360 365

Ala Met Leu Glu Leu Thr Gly Gln Asn Phe Thr Pro Asn Leu Arg Val

370 375 380

Trp Phe Gly Asp Val Glu Ala Glu Thr Met Tyr Arg Cys Gly Glu Ser

385 390 395 400

Met Leu Cys Val Val Pro Asp Ile Ser Ala Phe Arg Glu Gly Trp Arg

405 410 415

Trp Val Arg Gln Pro Val Gln Val Pro Val Thr Leu Val Arg Asn Asp

420 425 430

Gly Ile Ile Tyr Ser Thr Ser Leu Thr Phe Thr Tyr Thr Pro Glu Pro

435 440 445

Gly Pro Arg Pro His Cys Ser Ala Ala Gly Ala Ile Leu Arg Ala Asn

450 455 460

Ser Ser Gln Val Pro Pro Asn Glu Ser Asn Thr Asn Ser Glu Gly Ser

465 470 475 480

Tyr Thr Asn Ala Ser Thr Asn Ser Thr Ser Val Thr Ser Ser Thr Ala

485 490 495

Thr Val Val Ser

500

<210> 27

<211> 925

<212> DNA

<213> Intelligent people

<400> 27

acacacacat ctgcacctca accacagact acacttgctg aactggctcc tggggccatg 60

aggctgtcac tgccactgct gctgctgctg ctgggagcct gggccatccc agggggcctc 120

ggggacaggg cgccactcac agccacagcc ccacaactgg atgatgagga gatgtactca 180

gcccacatgc ccgctcacct gcgctgtgat gcctgcagag ctgtggctta ccagatgtgg 240

caaaatctgg caaaggcaga gaccaaactt catacctcaa actctggggg gcggcgggag 300

ctgagcgagt tggtctacac ggatgtcctg gaccggagct gctcccggaa ctggcaggac 360

tacggagttc gagaagtgga ccaagtgaaa cgtctcacag gcccaggact tagcgagggg 420

ccagagccaa gcatcagcgt gatggtcaca gggggcccct ggcctaccag gctctccagg 480

acatgtttgc actacttggg ggagtttgga gaagaccaga tctatgaagc ccaccaacaa 540

ggccgagggg ctctggaggc attgctatgt gggggacccc agggggcctg ctcagagaag 600

gtgtcagcca caagagaaga gctctagtcc tggactctac cctcctctga aagaagctgg 660

ggcttgctct gacggtctcc actcccgtct gcaggcagcc aggagggcag gaagcccttg 720

ctctgtgctg ccatcctgcc tccctcctcc agcctcaggg cactcgggcc tgggtgggag 780

tcaacgcctt cccctctgga ctcaaataaa acccagtgac ctcacttctt ttctctgcaa 840

aaggtgcttg tggggctggg agtgcagaca ttggtgtttc tgctgatgtc ccttgtgaaa 900

aaaaaaaaaa aactctcaaa atcaa 925

<210> 28

<211> 189

<212> PRT

<213> Intelligent people

<400> 28

Met Arg Leu Ser Leu Pro Leu Leu Leu Leu Leu Leu Gly Ala Trp Ala

1 5 10 15

Ile Pro Gly Gly Leu Gly Asp Arg Ala Pro Leu Thr Ala Thr Ala Pro

20 25 30

Gln Leu Asp Asp Glu Glu Met Tyr Ser Ala His Met Pro Ala His Leu

35 40 45

Arg Cys Asp Ala Cys Arg Ala Val Ala Tyr Gln Met Trp Gln Asn Leu

50 55 60

Ala Lys Ala Glu Thr Lys Leu His Thr Ser Asn Ser Gly Gly Arg Arg

65 70 75 80

Glu Leu Ser Glu Leu Val Tyr Thr Asp Val Leu Asp Arg Ser Cys Ser

85 90 95

Arg Asn Trp Gln Asp Tyr Gly Val Arg Glu Val Asp Gln Val Lys Arg

100 105 110

Leu Thr Gly Pro Gly Leu Ser Glu Gly Pro Glu Pro Ser Ile Ser Val

115 120 125

Met Val Thr Gly Gly Pro Trp Pro Thr Arg Leu Ser Arg Thr Cys Leu

130 135 140

His Tyr Leu Gly Glu Phe Gly Glu Asp Gln Ile Tyr Glu Ala His Gln

145 150 155 160

Gln Gly Arg Gly Ala Leu Glu Ala Leu Leu Cys Gly Gly Pro Gln Gly

165 170 175

Ala Cys Ser Glu Lys Val Ser Ala Thr Arg Glu Glu Leu

180 185

<210> 29

<211> 741

<212> DNA

<213> Intelligent people

<400> 29

gagaggctga gactaaccca gaaacatcca attctcaaac tgaagctcgc actctcgcct 60

ccagcatgaa agtctctgcc gcccttctgt gcctgctgct catagcagcc accttcattc 120

cccaagggct cgctcagcca gatgcaatca atgccccagt cacctgctgt tataacttca 180

ccaataggaa gatctcagtg cagaggctcg cgagctatag aagaatcacc agcagcaagt 240

gtcccaaaga agctgtgatc ttcaagacca ttgtggccaa ggagatctgt gctgacccca 300

agcagaagtg ggttcaggat tccatggacc acctggacaa gcaaacccaa actccgaaga 360

cttgaacact cactccacaa cccaagaatc tgcagctaac ttattttccc ctagctttcc 420

ccagacaccc tgttttattt tattataatg aattttgttt gttgatgtga aacattatgc 480

cttaagtaat gttaattctt atttaagtta ttgatgtttt aagtttatct ttcatggtac 540

tagtgttttt tagatacaga gacttgggga aattgctttt cctcttgaac cacagttcta 600

cccctgggat gttttgaggg tctttgcaag aatcattaat acaaagaatt ttttttaaca 660

ttccaatgca ttgctaaaat attattgtgg aaatgaatat tttgtaacta ttacaccaaa 720

taaatatatt tttgtacaaa a 741

<210> 30

<211> 99

<212> PRT

<213> Intelligent people

<400> 30

Met Lys Val Ser Ala Ala Leu Leu Cys Leu Leu Leu Ile Ala Ala Thr

1 5 10 15

Phe Ile Pro Gln Gly Leu Ala Gln Pro Asp Ala Ile Asn Ala Pro Val

20 25 30

Thr Cys Cys Tyr Asn Phe Thr Asn Arg Lys Ile Ser Val Gln Arg Leu

35 40 45

Ala Ser Tyr Arg Arg Ile Thr Ser Ser Lys Cys Pro Lys Glu Ala Val

50 55 60

Ile Phe Lys Thr Ile Val Ala Lys Glu Ile Cys Ala Asp Pro Lys Gln

65 70 75 80

Lys Trp Val Gln Asp Ser Met Asp His Leu Asp Lys Gln Thr Gln Thr

85 90 95

Pro Lys Thr

<210> 31

<211> 780

<212> DNA

<213> Intelligent people

<400> 31

agaaggacac gggcagcaga cagtggtcag tcctttcttg gctctgctga cactcgagcc 60

cacattccgt cacctgctca gaatcatgca ggtctccact gctgcccttg ctgtcctcct 120

ctgcaccatg gctctctgca accagttctc tgcatcactt gctgctgaca cgccgaccgc 180

ctgctgcttc agctacacct cccggcagat tccacagaat ttcatagctg actactttga 240

gacgagcagc cagtgctcca agcccggtgt catcttccta accaagcgaa gccggcaggt 300

ctgtgctgac cccagtgagg agtgggtcca gaaatatgtc agcgacctgg agctgagtgc 360

ctgaggggtc cagaagcttc gaggcccagc gacctcggtg ggcccagtgg ggaggagcag 420

gagcctgagc cttgggaaca tgcgtgtgac ctccacagct acctcttcta tggactggtt 480

gttgccaaac agccacactg tgggactctt cttaacttaa attttaattt atttatacta 540

tttagttttt gtaatttatt ttcgatttca cagtgtgttt gtgattgttt gctctgagag 600

ttcccctgtc ccctccccct tccctcacac cgcgtctggt gacaaccgag tggctgtcat 660

cagcctgtgt aggcagtcat ggcaccaaag ccaccagact gacaaatgtg tatcggatgc 720

ttttgttcag ggctgtgatc ggcctgggga aataataaag atgctctttt aaaaggtaaa 780

<210> 32

<211> 92

<212> PRT

<213> Intelligent people

<400> 32

Met Gln Val Ser Thr Ala Ala Leu Ala Val Leu Leu Cys Thr Met Ala

1 5 10 15

Leu Cys Asn Gln Phe Ser Ala Ser Leu Ala Ala Asp Thr Pro Thr Ala

20 25 30

Cys Cys Phe Ser Tyr Thr Ser Arg Gln Ile Pro Gln Asn Phe Ile Ala

35 40 45

Asp Tyr Phe Glu Thr Ser Ser Gln Cys Ser Lys Pro Gly Val Ile Phe

50 55 60

Leu Thr Lys Arg Ser Arg Gln Val Cys Ala Asp Pro Ser Glu Glu Trp

65 70 75 80

Val Gln Lys Tyr Val Ser Asp Leu Glu Leu Ser Ala

85 90

<210> 33

<211> 660

<212> DNA

<213> Intelligent people

<400> 33

agcacaggac acagctgggt tctgaagctt ctgagttctg cagcctcacc tctgagaaaa 60

cctcttttcc accaatacca tgaagctctg cgtgactgtc ctgtctctcc tcatgctagt 120

agctgccttc tgctctccag cgctctcagc accaatgggc tcagaccctc ccaccgcctg 180

ctgcttttct tacaccgcga ggaagcttcc tcgcaacttt gtggtagatt actatgagac 240

cagcagcctc tgctcccagc cagctgtggt attccaaacc aaaagaagca agcaagtctg 300

tgctgatccc agtgaatcct gggtccagga gtacgtgtat gacctggaac tgaactgagc 360

tgctcagaga caggaagtct tcagggaagg tcacctgagc ccggatgctt ctccatgaga 420

cacatctcct ccatactcag gactcctctc cgcagttcct gtcccttctc ttaatttaat 480

cttttttatg tgccgtgtta ttgtattagg tgtcatttcc attatttata ttagtttagc 540

caaaggataa gtgtccccta tggggatggt ccactgtcac tgtttctctg ctgttgcaaa 600

tacatggata acacatttga ttctgtgtgt tttcataata aaactttaaa ataaaatgca 660

<210> 34

<211> 92

<212> PRT

<213> Intelligent people

<400> 34

Met Lys Leu Cys Val Thr Val Leu Ser Leu Leu Met Leu Val Ala Ala

1 5 10 15

Phe Cys Ser Pro Ala Leu Ser Ala Pro Met Gly Ser Asp Pro Pro Thr

20 25 30

Ala Cys Cys Phe Ser Tyr Thr Ala Arg Lys Leu Pro Arg Asn Phe Val

35 40 45

Val Asp Tyr Tyr Glu Thr Ser Ser Leu Cys Ser Gln Pro Ala Val Val

50 55 60

Phe Gln Thr Lys Arg Ser Lys Gln Val Cys Ala Asp Pro Ser Glu Ser

65 70 75 80

Trp Val Gln Glu Tyr Val Tyr Asp Leu Glu Leu Asn

85 90

<210> 35

<211> 309

<212> DNA

<213> Intelligent people

<400> 35

accatgaagg tctccgcggc agccctcgct gtcatcctca ttgctactgc cctctgcgct 60

cctgcatctg cctccccata ttcctcggac accacaccct gctgctttgc ctacattgcc 120

cgcccactgc cccgtgccca catcaaggag tatttctaca ccagtggcaa gtgctccaac 180

ccagcagtcg tctttgtcac ccgaaagaac cgccaagtgt gtgccaaccc agagaagaaa 240

tgggttcggg agtacatcaa ctctttggag atgagctagg atggagagtc cttgaacctg 300

aacttacac 309

<210> 36

<211> 91

<212> PRT

<213> Intelligent

<400> 36

Met Lys Val Ser Ala Ala Ala Leu Ala Val Ile Leu Ile Ala Thr Ala

1 5 10 15

Leu Cys Ala Pro Ala Ser Ala Ser Pro Tyr Ser Ser Asp Thr Thr Pro

20 25 30

Cys Cys Phe Ala Tyr Ile Ala Arg Pro Leu Pro Arg Ala His Ile Lys

35 40 45

Glu Tyr Phe Tyr Thr Ser Gly Lys Cys Ser Asn Pro Ala Val Val Phe

50 55 60

Val Thr Arg Lys Asn Arg Gln Val Cys Ala Asn Pro Glu Lys Lys Trp

65 70 75 80

Val Arg Glu Tyr Ile Asn Ser Leu Glu Met Ser

85 90

<210> 37

<211> 862

<212> DNA

<213> Intelligent

<400> 37

agagaggttg agaacaaccc agaaaccttc acctctcatg ctgaagctca cacccttgcc 60

ctccaagatg aaggtttctg cagcgcttct gtgcctgctg ctcatggcag ccactttcag 120

ccctcaggga cttgctcagc cagattcagt ttccattcca atcacctgct gctttaacgt 180

gatcaatagg aaaattccta tccagaggct ggagagctac acaagaatca ccaacatcca 240

atgtcccaag gaagctgtga tcttcaagac caaacggggc aaggaggtct gtgctgaccc 300

caaggagaga tgggtcaggg attccatgaa gcatctggac caaatatttc aaaatctgaa 360

gccatgagcc ttcatacatg gactgagagt cagagcttga agaaaagctt atttattttc 420

cccaacctcc cccaggtgca gtgtgacatt attttattat aacatccaca aagagattat 480

ttttaaataa tttaaagcat aatatttctt aaaaagtatt taattatatt taagttgttg 540

atgttttaac tctatctgtc atacatccta gtgaatgtaa aatgcaaaat cctggtgatg 600

tgttttttgt ttttgttttc ctgtgagctc aactaagttc acggcaaaat gtcattgttc 660

tccctcctac ctgtctgtag tgttgtgggg tcctcccatg gatcatcaag gtgaaacact 720

ttggtattct ttggcaatca gtgctcctgt aagtcaaatg tgtgctttgt actgctgttg 780

ttgaaattga tgttactgta tataactatg gaattttgaa aaaaaatttc aaaaagaaaa 840

aaatatatat aatttaaaac ta 862

<210> 38

<211> 99

<212> PRT

<213> Intelligent people

<400> 38

Met Lys Val Ser Ala Ala Leu Leu Cys Leu Leu Leu Met Ala Ala Thr

1 5 10 15

Phe Ser Pro Gln Gly Leu Ala Gln Pro Asp Ser Val Ser Ile Pro Ile

20 25 30

Thr Cys Cys Phe Asn Val Ile Asn Arg Lys Ile Pro Ile Gln Arg Leu

35 40 45

Glu Ser Tyr Thr Arg Ile Thr Asn Ile Gln Cys Pro Lys Glu Ala Val

50 55 60

Ile Phe Lys Thr Lys Arg Gly Lys Glu Val Cys Ala Asp Pro Lys Glu

65 70 75 80

Arg Trp Val Arg Asp Ser Met Lys His Leu Asp Gln Ile Phe Gln Asn

85 90 95

Leu Lys Pro

<210> 39

<211> 1332

<212> DNA

<213> Intelligent

<400> 39

agaaggaggc caggagttgt gagtttccaa gccccagctc actctgacca cttctctgcc 60

tgcccagcat catgaagggc cttgcagctg ccctccttgt cctcgtctgc accatggccc 120

tctgctcctg tgcacaagtt ggtaccaaca aagagctctg ctgcctcgtc tatacctcct 180

ggcagattcc acaaaagttc atagttgact attctgaaac cagcccccag tgccccaagc 240

caggtgtcat cctcctaacc aagagaggcc ggcagatctg tgctgacccc aataagaagt 300

gggtccagaa atacatcagc gacctgaagc tgaatgcctg aggggcctgg aagctgcgag 360

ggcccagtga acttggtggg cccaggaggg aacaggagcc tgagccaggg caatggccct 420

gccaccctgg aggctacctc ttctaagagt cccatctgct atgcccagcc acattaacta 480

actttaatct tagtttatgc atcatatttc attttgaaat tgatttctat tgttgagctg 540

cattatgaaa ttagtatttt ctctgacatc tcatgacatt gtctttatca tcctttcccc 600

tttcccttca actcttcgta cattcaatgc atggatcaat cagtgtgatt agctttctca 660

gcagacattg tgccatatgt atcaaatgac aaatctttat tgaatggttt tgctcagcac 720

caccttttaa tatattggca gtacttatta tataaaaggt aaaccagcat tctcactgtg 780

acgactctgt tgattttgtt tcactaatcg gaatcacaga ctgagaggaa ttctggggga 840

ggagtaggga aatatgaaaa ggggcagaaa cagcaagata gggaccaagt acttctacag 900

tcataccaga catttccctg gagatacttt cctgaaaagt tgaaacagac accattagtt 960

tataaaccat attgtaactg aaatgtgata gaaaaatttt ctacttaaat gaatatcaag 1020

atgacgctgc aatgcatatt tacgcacaca agctcatttt cataaatgaa gctttcctca 1080

gaatgatggc taacatctgt tgaggtctta ccaagtgctg ggttttgggc taagtactta 1140

tattagtaaa agtctagcaa atactgccca tagtctagca aggactcctt acctggaagt 1200

tgctgaaagc cttggtaatg ttatctttgt tctccactct gcttttgggg aggatgtttt 1260

ccatgactat acgagatgag gcttggggct gggtggccag agttagcaaa taaaaataca 1320

gaatggatac ta 1332

<210> 40

<211> 89

<212> PRT

<213> Intelligent people

<400> 40

Met Lys Gly Leu Ala Ala Ala Leu Leu Val Leu Val Cys Thr Met Ala

1 5 10 15

Leu Cys Ser Cys Ala Gln Val Gly Thr Asn Lys Glu Leu Cys Cys Leu

20 25 30

Val Tyr Thr Ser Trp Gln Ile Pro Gln Lys Phe Ile Val Asp Tyr Ser

35 40 45

Glu Thr Ser Pro Gln Cys Pro Lys Pro Gly Val Ile Leu Leu Thr Lys

50 55 60

Arg Gly Arg Gln Ile Cys Ala Asp Pro Asn Lys Lys Trp Val Gln Lys

65 70 75 80

Tyr Ile Ser Asp Leu Lys Leu Asn Ala

85

<210> 41

<211> 683

<212> DNA

<213> Intelligent

<400> 41

attcccagcc tcacatcact cacaccttgc atttcacccc tgcatcccag tcgccctgca 60

gcctcacaca gatcctgcac acacccagac agctggcgct cacacattca ccgttggcct 120

gcctctgttc accctccatg gccctgctac tggccctcag cctgctggtt ctctggactt 180

ccccagcccc aactctgagt ggcaccaatg atgctgaaga ctgctgcctg tctgtgaccc 240

agaaacccat ccctgggtac atcgtgagga acttccacta ccttctcatc aaggatggct 300

gcagggtgcc tgctgtagtg ttcaccacac tgaggggccg ccagctctgt gcacccccag 360

accagccctg ggtagaacgc atcatccaga gactgcagag gacctcagcc aagatgaagc 420

gccgcagcag ttaacctatg accgtgcaga gggagcccgg agtccgagtc aagcattgtg 480

aattattacc taacctgggg aaccgaggac cagaaggaag gaccaggctt ccagctcctc 540

tgcaccagac ctgaccagcc aggacagggc ctggggtgtg tgtgagtgtg agtgtgagcg 600

agagggtgag tgtggtcaga gtaaagctgc tccaccccca gattgcaatg ctaccaataa 660

agccgcctgg tgtttacaac taa 683

<210> 42

<211> 98

<212> PRT

<213> Intelligent

<400> 42

Met Ala Leu Leu Leu Ala Leu Ser Leu Leu Val Leu Trp Thr Ser Pro

1 5 10 15

Ala Pro Thr Leu Ser Gly Thr Asn Asp Ala Glu Asp Cys Cys Leu Ser

20 25 30

Val Thr Gln Lys Pro Ile Pro Gly Tyr Ile Val Arg Asn Phe His Tyr

35 40 45

Leu Leu Ile Lys Asp Gly Cys Arg Val Pro Ala Val Val Phe Thr Thr

50 55 60

Leu Arg Gly Arg Gln Leu Cys Ala Pro Pro Asp Gln Pro Trp Val Glu

65 70 75 80

Arg Ile Ile Gln Arg Leu Gln Arg Thr Ser Ala Lys Met Lys Arg Arg

85 90 95

Ser Ser

<210> 43

<211> 864

<212> DNA

<213> Intelligent people

<400> 43

acagaccccc aacttgcagc tgcccacctc accctcagct ctggcctctt actcaccctc 60

taccacagac atggctcagt cactggctct gagcctcctt atcctggttc tggcctttgg 120

catccccagg acccaaggca gtgatggagg ggctcaggac tgttgcctca agtacagcca 180

aaggaagatt cccgccaagg ttgtccgcag ctaccggaag caggaaccaa gcttaggctg 240

ctccatccca gctatcctgt tcttgccccg caagcgctct caggcagagc tatgtgcaga 300

cccaaaggag ctctgggtgc agcagctgat gcagcatctg gacaagacac catccccaca 360

gaaaccagcc cagggctgca ggaaggacag gggggcctcc aagactggca agaaaggaaa 420

gggctccaaa ggctgcaaga ggactgagcg gtcacagacc cctaaagggc catagcccag 480

tgagcagcct ggagccctgg agaccccacc agcctcacca gcgcttgaag cctgaaccca 540

agatgcaaga aggaggctat gctcaggggc cctggagcag ccaccccatg ctggccttgc 600

cacactcttt ctcctgcttt aaccacccca tctgcattcc cagctctacc ctgcatggct 660

gagctgccca cagcaggcca ggtccagaga gaccgaggag ggagagtctc ccagggagca 720

tgagaggagg cagcaggact gtccccttga aggagaatca tcaggaccct ggacctgata 780

cggctcccca gtacacccca cctcttcctt gtaaatatga tttataccta actgaataaa 840

aagctgttct gtcttcccac ccaa 864

<210> 44

<211> 134

<212> PRT

<213> Intelligent

<400> 44

Met Ala Gln Ser Leu Ala Leu Ser Leu Leu Ile Leu Val Leu Ala Phe

1 5 10 15

Gly Ile Pro Arg Thr Gln Gly Ser Asp Gly Gly Ala Gln Asp Cys Cys

20 25 30

Leu Lys Tyr Ser Gln Arg Lys Ile Pro Ala Lys Val Val Arg Ser Tyr

35 40 45

Arg Lys Gln Glu Pro Ser Leu Gly Cys Ser Ile Pro Ala Ile Leu Phe

50 55 60

Leu Pro Arg Lys Arg Ser Gln Ala Glu Leu Cys Ala Asp Pro Lys Glu

65 70 75 80

Leu Trp Val Gln Gln Leu Met Gln His Leu Asp Lys Thr Pro Ser Pro

85 90 95

Gln Lys Pro Ala Gln Gly Cys Arg Lys Asp Arg Gly Ala Ser Lys Thr

100 105 110

Gly Lys Lys Gly Lys Gly Ser Lys Gly Cys Lys Arg Thr Glu Arg Ser

115 120 125

Gln Thr Pro Lys Gly Pro

130

<210> 45

<211> 2761

<212> DNA

<213> Intelligent people

<400> 45

aaagaatttc tcaggctcaa aatccaatac aggagtgact tggaactcca ttctatcact 60

atgaagaaaa gtggtgttct tttcctcttg ggcatcatct tgctggttct gattggagtg 120

caaggaaccc cagtagtgag aaagggtcgc tgttcctgca tcagcaccaa ccaagggact 180

atccacctac aatccttgaa agaccttaaa caatttgccc caagcccttc ctgcgagaaa 240

attgaaatca ttgctacact gaagaatgga gttcaaacat gtctaaaccc agattcagca 300

gatgtgaagg aactgattaa aaagtgggag aaacaggtca gccaaaagaa aaagcaaaag 360

aatgggaaaa aacatcaaaa aaagaaagtt ctgaaagttc gaaaatctca acgttctcgt 420

caaaagaaga ctacataaga gaccacttca ccaataagta ttctgtgtta aaaatgttct 480

attttaatta taccgctatc attccaaagg aggatggcat ataatacaaa ggcttattaa 540

tttgactaga aaatttaaaa cattactctg aaattgtaac taaagttaga aagttgattt 600

taagaatcca aacgttaaga attgttaaag gctatgattg tctttgttct tctaccaccc 660

accagttgaa tttcatcatg cttaaggcca tgattttagc aatacccatg tctacacaga 720

tgttcaccca accacatccc actcacaaca gctgcctgga agagcagccc taggcttcca 780

cgtactgcag cctccagaga gtatctgagg cacatgtcag caagtcctaa gcctgttagc 840

atgctggtga gccaagcagt ttgaaattga gctggacctc accaagctgc tgtggccatc 900

aacctctgta tttgaatcag cctacaggcc tcacacacaa tgtgtctgag agattcatgc 960

tgattgttat tgggtatcac cactggagat caccagtgtg tggctttcag agcctccttt 1020

ctggctttgg aagccatgtg attccatctt gcccgctcag gctgaccact ttatttcttt 1080

ttgttcccct ttgcttcatt caagtcagct cttctccatc ctaccacaat gcagtgcctt 1140

tcttctctcc agtgcacctg tcatatgctc tgatttatct gagtcaactc ctttctcatc 1200

ttgtccccaa caccccacag aagtgctttc ttctcccaat tcatcctcac tcagtccagc 1260

ttagttcaag tcctgcctct taaataaacc tttttggaca cacaaattat cttaaaactc 1320

ctgtttcact tggttcagta ccacatgggt gaacactcaa tggttaacta attcttgggt 1380

gtttatccta tctctccaac cagattgtca gctccttgag ggcaagagcc acagtatatt 1440

tccctgtttc ttccacagtg cctaataata ctgtggaact aggttttaat aattttttaa 1500

ttgatgttgt tatgggcagg atggcaacca gaccattgtc tcagagcagg tgctggctct 1560

ttcctggcta ctccatgttg gctagcctct ggtaacctct tacttattat cttcaggaca 1620

ctcactacag ggaccaggga tgatgcaaca tccttgtctt tttatgacag gatgtttgct 1680

cagcttctcc aacaataaga agcacgtggt aaaacacttg cggatattct ggactgtttt 1740

taaaaaatat acagtttacc gaaaatcata taatcttaca atgaaaagga ctttatagat 1800

cagccagtga ccaacctttt cccaaccata caaaaattcc ttttcccgaa ggaaaagggc 1860

tttctcaata agcctcagct ttctaagatc taacaagata gccaccgaga tccttatcga 1920

aactcatttt aggcaaatat gagttttatt gtccgtttac ttgtttcaga gtttgtattg 1980

tgattatcaa ttaccacacc atctcccatg aagaaaggga acggtgaagt actaagcgct 2040

agaggaagca gccaagtcgg ttagtggaag catgattggt gcccagttag cctctgcagg 2100

atgtggaaac ctccttccag gggaggttca gtgaattgtg taggagaggt tgtctgtggc 2160

cagaatttaa acctatactc actttcccaa attgaatcac tgctcacact gctgatgatt 2220

tagagtgctg tccggtggag atcccacccg aacgtcttat ctaatcatga aactccctag 2280

ttccttcatg taacttccct gaaaaatcta agtgtttcat aaatttgaga gtctgtgacc 2340

cacttacctt gcatctcaca ggtagacagt atataactaa caaccaaaga ctacatattg 2400

tcactgacac acacgttata atcatttatc atatatatac atacatgcat acactctcaa 2460

agcaaataat ttttcacttc aaaacagtat tgacttgtat accttgtaat ttgaaatatt 2520

ttctttgtta aaatagaatg gtatcaataa atagaccatt aatcagaaaa cagatcttga 2580

ttttttttct cttgaatgta cccttcaact gttgaatgtt taatagtaaa tcttatatgt 2640

ccttatttac tttttagctt tctctcaaat aaagtgtaac actagttgag ataacacatg 2700

aaagctcttt aaagggtcga tcgggaacag gaaaaaaaac ctatggaaaa tatgacaaca 2760

c 2761

<210> 46

<211> 125

<212> PRT

<213> Intelligent people

<400> 46

Met Lys Lys Ser Gly Val Leu Phe Leu Leu Gly Ile Ile Leu Leu Val

1 5 10 15

Leu Ile Gly Val Gln Gly Thr Pro Val Val Arg Lys Gly Arg Cys Ser

20 25 30

Cys Ile Ser Thr Asn Gln Gly Thr Ile His Leu Gln Ser Leu Lys Asp

35 40 45

Leu Lys Gln Phe Ala Pro Ser Pro Ser Cys Glu Lys Ile Glu Ile Ile

50 55 60

Ala Thr Leu Lys Asn Gly Val Gln Thr Cys Leu Asn Pro Asp Ser Ala

65 70 75 80

Asp Val Lys Glu Leu Ile Lys Lys Trp Glu Lys Gln Val Ser Gln Lys

85 90 95

Lys Lys Gln Lys Asn Gly Lys Lys His Gln Lys Lys Lys Val Leu Lys

100 105 110

Val Arg Lys Ser Gln Arg Ser Arg Gln Lys Lys Thr Thr

115 120 125

<210> 47

<211> 1175

<212> DNA

<213> Intelligent

<400> 47

gagacattcc tcaattgctt agacatattc tgagcctaca gcagaggaac ctccagtctc 60

agcaccatga atcaaactgc cattctgatt tgctgcctta tctttctgac tctaagtggc 120

attcaaggag tacctctctc tagaactgta cgctgtacct gcatcagcat tagtaatcaa 180

cctgttaatc caaggtcttt agaaaaactt gaaattattc ctgcaagcca attttgtcca 240

cgtgttgaga tcattgctac aatgaaaaag aagggtgaga agagatgtct gaatccagaa 300

tcgaaggcca tcaagaattt actgaaagca gttagcaagg aaaggtctaa aagatctcct 360

taaaaccaga ggggagcaaa atcgatgcag tgcttccaag gatggaccac acagaggctg 420

cctctcccat cacttcccta catggagtat atgtcaagcc ataattgttc ttagtttgca 480

gttacactaa aaggtgacca atgatggtca ccaaatcagc tgctactact cctgtaggaa 540

ggttaatgtt catcatccta agctattcag taataactct accctggcac tataatgtaa 600

gctctactga ggtgctatgt tcttagtgga tgttctgacc ctgcttcaaa tatttccctc 660

acctttccca tcttccaagg gtactaagga atctttctgc tttggggttt atcagaattc 720

tcagaatctc aaataactaa aaggtatgca atcaaatctg ctttttaaag aatgctcttt 780

acttcatgga cttccactgc catcctccca aggggcccaa attctttcag tggctaccta 840

catacaattc caaacacata caggaaggta gaaatatctg aaaatgtatg tgtaagtatt 900

cttatttaat gaaagactgt acaaagtaga agtcttagat gtatatattt cctatattgt 960

tttcagtgta catggaataa catgtaatta agtactatgt atcaatgagt aacaggaaaa 1020

ttttaaaaat acagatagat atatgctctg catgttacat aagataaatg tgctgaatgg 1080

ttttcaaaat aaaaatgagg tactctcctg gaaatattaa gaaagactat ctaaatgttg 1140

aaagatcaaa aggttaataa agtaattata actaa 1175

<210> 48

<211> 98

<212> PRT

<213> Intelligent

<400> 48

Met Asn Gln Thr Ala Ile Leu Ile Cys Cys Leu Ile Phe Leu Thr Leu

1 5 10 15

Ser Gly Ile Gln Gly Val Pro Leu Ser Arg Thr Val Arg Cys Thr Cys

20 25 30

Ile Ser Ile Ser Asn Gln Pro Val Asn Pro Arg Ser Leu Glu Lys Leu

35 40 45

Glu Ile Ile Pro Ala Ser Gln Phe Cys Pro Arg Val Glu Ile Ile Ala

50 55 60

Thr Met Lys Lys Lys Gly Glu Lys Arg Cys Leu Asn Pro Glu Ser Lys

65 70 75 80

Ala Ile Lys Asn Leu Leu Lys Ala Val Ser Lys Glu Arg Ser Lys Arg

85 90 95

Ser Pro

<210> 49

<211> 1479

<212> DNA

<213> Intelligent people

<400> 49

gttcagcatt tctactcctt ccaagaagag cagcaaagct gaagtagcag cagcagcacc 60

agcagcaaca gcaaaaaaca aacatgagtg tgaagggcat ggctatagcc ttggctgtga 120

tattgtgtgc tacagttgtt caaggcttcc ccatgttcaa aagaggacgc tgtctttgca 180

taggccctgg ggtaaaagca gtgaaagtgg cagatattga gaaagcctcc ataatgtacc 240

caagtaacaa ctgtgacaaa atagaagtga ttattaccct gaaagaaaat aaaggacaac 300

gatgcctaaa tcccaaatcg aagcaagcaa ggcttataat caaaaaagtt gaaagaaaga 360

atttttaaaa atatcaaaac atatgaagtc ctggaaaaga gcatctgaaa aacctagaac 420

aagtttaact gtgactactg aaatgacaag aattctacag taggaaactg agacttttct 480

atggttttgt gactttcaac ttttgtacag ttatgtgaag gatgaaaggt gggtgaaagg 540

accaaaaaca gaaatacagt cttcctgaat gaatgacaat cagaattcca ctgcccaaag 600

gagtccaaca attaaatgga tttctaggaa aagctacctt aagaaaggct ggttaccatc 660

ggagtttaca aagtgctttc acgttcttac ttgttgcatt atacattcat gcatttctag 720

gctagagaac cttctagatt tgatgcttac aactattctg ttgtgactat gagaacattt 780

ctgtctctag aagtcatctg tctgtattga tctttatgct atattactat ctgtggttac 840

ggtggagaca ttgacattat tactggagtc aagcccttat aagtcaaaag catctatgtg 900

tcgtaaaaca ttcctcaaac attttttcat gcaaatacac acttctttcc ccaaacatca 960

tgtagcacat caatatgtag ggagacattc ttatgcatca tttggtttgt tttataacca 1020

attcattaaa tgtaattcat aaaatgtact atgaaaaaaa ttatacgcta tgggatactg 1080

gcaaaagtgc acatatttca taaccaaatt agtagcacca gtcttaattt gatgtttttc 1140

aacttttatt cattgagatg ttttgaagca attaggatat gtgtgtttac tgtacttttt 1200

gttttgatcc gtttgtataa atgatagcaa tatcttggac acatctgaaa tacaaaatgt 1260

ttttgtctac caaagaaaaa tgttgaaaaa taagcaaatg tatacctagc aatcactttt 1320

actttttgta attctgtctc ttagaaaaat acataatcta atcaatttct ttgttcatgc 1380

ctatatactg taaaatttag gtatactcaa gactagttta aagaatcaaa gtcatttttt 1440

tctctaataa actaccacaa cctttctttt ttaaaaaaa 1479

<210> 50

<211> 94

<212> PRT

<213> Intelligent people

<400> 50

Met Ser Val Lys Gly Met Ala Ile Ala Leu Ala Val Ile Leu Cys Ala

1 5 10 15

Thr Val Val Gln Gly Phe Pro Met Phe Lys Arg Gly Arg Cys Leu Cys

20 25 30

Ile Gly Pro Gly Val Lys Ala Val Lys Val Ala Asp Ile Glu Lys Ala

35 40 45

Ser Ile Met Tyr Pro Ser Asn Asn Cys Asp Lys Ile Glu Val Ile Ile

50 55 60

Thr Leu Lys Glu Asn Lys Gly Gln Arg Cys Leu Asn Pro Lys Ser Lys

65 70 75 80

Gln Ala Arg Leu Ile Ile Lys Lys Val Glu Arg Lys Asn Phe

85 90

<210> 51

<211> 1219

<212> DNA

<213> Intelligent people

<400> 51

gagaagatgt ttgaaaaaac tgactctgct aatgagcctg gactcagagc tcaagtctga 60

actctacctc cagacagaat gaagttcatc tcgacatctc tgcttctcat gctgctggtc 120

agcagcctct ctccagtcca aggtgttctg gaggtctatt acacaagctt gaggtgtaga 180

tgtgtccaag agagctcagt ctttatccct agacgcttca ttgatcgaat tcaaatcttg 240

ccccgtggga atggttgtcc aagaaaagaa atcatagtct ggaagaagaa caagtcaatt 300

gtgtgtgtgg accctcaagc tgaatggata caaagaatga tggaagtatt gagaaaaaga 360

agttcttcaa ctctaccagt tccagtgttt aagagaaaga ttccctgatg ctgatatttc 420

cactaagaac acctgcattc ttcccttatc cctgctctgg attttagttt tgtgcttagt 480

taaatctttt ccaggaaaaa gaacttcccc atacaaataa gcatgagact atgtaaaaat 540

aaccttgcag aagctgatgg ggcaaactca agcttcttca ctcacagcac cctatataca 600

cttggagttt gcattcttat tcatcaggga ggaaagtttc tttgaaaata gttattcagt 660

tataagtaat acaggattat tttgattata tacttgttgt ttaatgttta aaatttctta 720

gaaaacaatg gaatgagaat ttaagcctca aatttgaaca tgtggcttga attaagaaga 780

aaattatggc atatattaaa agcaggcttc tatgaaagac tcaaaaagct gcctgggagg 840

cagatggaac ttgagcctgt caagaggcaa aggaatccat gtagtagata tcctctgctt 900

aaaaactcac tacggaggag aattaagtcc tacttttaaa gaatttcttt ataaaattta 960

ctgtctaaga ttaatagcat tcgaagatcc ccagacttca tagaatactc agggaaagca 1020

tttaaagggt gatgtacaca tgtatccttt cacacatttg ccttgacaaa cttctttcac 1080

tcacatcttt ttcactgact ttttttgtgg ggggcggggc cggggggact ctggtatcta 1140

attctttaat gattcctata aatctaatga cattcaataa agttgagcaa acattttact 1200

taaaaaaaaa aaaaaaaaa 1219

<210> 52

<211> 109

<212> PRT

<213> Intelligent people

<400> 52

Met Lys Phe Ile Ser Thr Ser Leu Leu Leu Met Leu Leu Val Ser Ser

1 5 10 15

Leu Ser Pro Val Gln Gly Val Leu Glu Val Tyr Tyr Thr Ser Leu Arg

20 25 30

Cys Arg Cys Val Gln Glu Ser Ser Val Phe Ile Pro Arg Arg Phe Ile

35 40 45

Asp Arg Ile Gln Ile Leu Pro Arg Gly Asn Gly Cys Pro Arg Lys Glu

50 55 60

Ile Ile Val Trp Lys Lys Asn Lys Ser Ile Val Cys Val Asp Pro Gln

65 70 75 80

Ala Glu Trp Ile Gln Arg Met Met Glu Val Leu Arg Lys Arg Ser Ser

85 90 95

Ser Thr Leu Pro Val Pro Val Phe Lys Arg Lys Ile Pro

100 105

<210> 53

<211> 1060

<212> DNA

<213> Intelligent people

<400> 53

cggctcccgc gccgcctccc ctcgcgcccg agcttcgagc caagcagcgt cctggggagc 60

gcgtcatggc cttaccagtg accgccttgc tcctgccgct ggccttgctg ctccacgccg 120

ccaggccgag ccagttccgg gtgtcgccgc tggatcggac ctggaacctg ggcgagacag 180

tggagctgaa gtgccaggtg ctgctgtcca acccgacgtc gggctgctcg tggctcttcc 240

agccgcgcgg cgccgccgcc agtcccacct tcctcctata cctctcccaa aacaagccca 300

aggcggccga ggggctggac acccagcggt tctcgggcaa gaggttgggg gacaccttcg 360

tcctcaccct gagcgacttc cgccgagaga acgagggcta ctatttctgc tcggccctga 420

gcaactccat catgtacttc agccacttcg tgccggtctt cctgccagcg aagcccacca 480

cgacgccagc gccgcgacca ccaacaccgg cgcccaccat cgcgtcgcag cccctgtccc 540

tgcgcccaga ggcgtgccgg ccagcggcgg ggggcgcagt gcacacgagg gggctggact 600

tcgcctgtga tatctacatc tgggcgccct tggccgggac ttgtggggtc cttctcctgt 660

cactggttat caccctttac tgcaaccaca ggaaccgaag acgtgtttgc aaatgtcccc 720

ggcctgtggt caaatcggga gacaagccca gcctttcggc gagatacgtc taaccctgtg 780

caacagccac tacattactt caaactgaga tccttccttt tgagggagca agtccttccc 840

tttcattttt tccagtcttc ctccctgtgt attcattctc atgattatta ttttagtggg 900

ggcggggtgg gaaagattac tttttcttta tgtgtttgac gggaaacaaa actaggtaaa 960

atctacagta caccacaagg gtcacaatac tgttgtgcgc acatcgcggt agggcgtgga 1020

aaggggcagg ccagagctac ccgcagagtt ctcagaatca 1060

<210> 54

<211> 235

<212> PRT

<213> Intelligent

<400> 54

Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu

1 5 10 15

His Ala Ala Arg Pro Ser Gln Phe Arg Val Ser Pro Leu Asp Arg Thr

20 25 30

Trp Asn Leu Gly Glu Thr Val Glu Leu Lys Cys Gln Val Leu Leu Ser

35 40 45

Asn Pro Thr Ser Gly Cys Ser Trp Leu Phe Gln Pro Arg Gly Ala Ala

50 55 60

Ala Ser Pro Thr Phe Leu Leu Tyr Leu Ser Gln Asn Lys Pro Lys Ala

65 70 75 80

Ala Glu Gly Leu Asp Thr Gln Arg Phe Ser Gly Lys Arg Leu Gly Asp

85 90 95

Thr Phe Val Leu Thr Leu Ser Asp Phe Arg Arg Glu Asn Glu Gly Tyr

100 105 110

Tyr Phe Cys Ser Ala Leu Ser Asn Ser Ile Met Tyr Phe Ser His Phe

115 120 125

Val Pro Val Phe Leu Pro Ala Lys Pro Thr Thr Thr Pro Ala Pro Arg

130 135 140

Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg

145 150 155 160

Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly

165 170 175

Leu Asp Phe Ala Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr

180 185 190

Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Asn His

195 200 205

Arg Asn Arg Arg Arg Val Cys Lys Cys Pro Arg Pro Val Val Lys Ser

210 215 220

Gly Asp Lys Pro Ser Leu Ser Ala Arg Tyr Val

225 230 235

<210> 55

<211> 3207

<212> DNA

<213> Intelligent people

<400> 55

agccagctgc cgtctgcgct ttgcaaagca tgcagttagg ggagcagctc ttggtgagct 60

cagtgaacct gcctggcgcg cacttctacc cgctggagag tgcgcgaggc ggcagcggcg 120

ggagcgctgg ccacctcccc agcgcggccc cctctcctca gaagttggac ttagacaaag 180

cgtccaagaa gttttccggc agtctctcct gcgaggcggt gagcggggag cccgcagccg 240

ccagcgcagg ggcccccgcg gccatgctta gtgacaccga cgccggggac gcatttgcca 300

gcgctgcggc agtggccaag ccggggcccc cggacggccg caagggctcc ccctgcgggg 360

aggaggagct gccctccgcc gctgcagccg ccgccgccgc cgccgccgcg gctgcggcca 420

ctgcgcgcta ctccatggac agcctgagct ccgagcggta ctacctccag tcccccggtc 480

ctcaggggtc ggagctggct gcgccctgct cactcttccc gtaccaggcg gcggctgggg 540

cgccccacgg acctgtgtac ccggctccta acggggcgcg ctacccctac ggctccatgc 600

tgccccccgg cggcttcccc gcggctgtgt gcccacccgg gagggcgcag ttcggcccag 660

gagccggtgc gggcagtggc gcgggcggta gcagcggcgg gggcggcggc ccgggcacct 720

atcagtacag ccagggggct ccgctctacg ggccgtaccc tggagccgca gcggcgggat 780

cttgcggagg actggggggc ctgggggttc caggttctgg cttccgtgcc cacgtctacc 840

tgtgcaaccg gcctctgtgg ctcaaattcc accgccacca aactgagatg atcattacga 900

aacagggcag gcgcatgttt cctttcttga gcttcaacat aaacggactc aatcccactg 960

cccactacaa tgtgttcgta gaggtggtgc tggcggaccc caaccactgg cgcttccagg 1020

ggggcaaatg ggtgacctgt ggcaaagccg acaataacat gcagggcaac aaaatgtatg 1080

ttcacccaga gtctcctaat actggttccc actggatgag acaggagatt tcattcggga 1140

aattaaaact caccaataac aaaggcgcaa ataacaacaa cacccagatg atagtcttac 1200

aatccttaca caaataccaa ccccgactgc atattgttga agttacagag gatggcgtgg 1260

aggacttgaa tgagccctca aagacccaga cttttacctt ctcagaaacg caattcattg 1320

cagtgactgc ctaccaaaac accgatatta ctcaactaaa gattgatcat aacccctttg 1380

caaaaggctt cagagacaac tatgattcat cccatcagat tgtccctgga ggtcggtacg 1440

gcgttcaatc cttcttcccg gagccctttg tcaacacttt acctcaagcc cgctattata 1500

atggcgagag aaccgtgcca cagaccaacg gcctcctttc accccaacag agcgaagagg 1560

tggccaaccc tccccagcgg tggcttgtca cgcctgtcca gcaacctggg accaacaaac 1620

tagacatcag ttcctatgaa tctgaatata cttctagcac attgctccca tatggcatta 1680

aatccttgcc ccttcagaca tcccatgccc tggggtatta cccagaccca acctttcctg 1740

caatggcagg gtggggaggt cgaggttctt accagaggaa gatggcagct ggactaccat 1800

ggacctccag aacaagcccc actgtgttct ctgaagatca gctctccaag gagaaagtga 1860

aagaggaaat tggctcttct tggatagaga cacccccttc catcaaatct ctagattcca 1920

atgattcagg agtatacacc agtgcttgta agcgaaggcg gctgtctcct agcaactcca 1980

gtaatgaaaa ttcaccctcc ataaagtgtg aggacattaa tgctgaagag tatagtaaag 2040

acacctcaaa aggcatggga gggtattatg ctttttacac aactccctaa agagttattt 2100

taacctcaaa aattagctaa ctttttgcag atggacttgg tggtgttttt tgttgtcttc 2160

tttgcctagg ttgccaaaaa gatgtttgcc ttccaccttg atgcatcctg ttttgtgcaa 2220

ttctctaaaa gaaggtgcca aagctttttg attgctgcag gtaactgaaa caaacctagc 2280

atttttaaaa aataagatta atggaagact ttaaggtatt ttaaaattcg aagggtatcc 2340

aaggttctgt atttatttat tggggagaca ctaacccttc aaagaagcag gctgtgaaca 2400

ttgggtgccc agtgctatca gatgagttaa aacctttgat tctcatttct atttgtaaat 2460

tcttaagcaa atagaagccg agtgttaagg tgttttgctt ctgaaagagg gctgtgcctt 2520

ccgtttcaga aggagacatt ttgctgttac attctgccag gggcaaaaga tactaggccc 2580

aggagtcaag aaaagctttt gtgaaagtga tagtttcacc tgactttgat tccttaaccc 2640

ccggcttttg gaacaagcca tgtttgccct agtccaggat tgcctcactt gagacttgct 2700

aggcctctgc tgtgtgctgg ggtggccagt gggactcagg agagagcaag ctaaggagtc 2760

accaaaaaaa aaaaaaaaaa aaagggagaa tttaaaagtg tacagttgtg tgtttagata 2820

cactatagaa taatgtggta tatattgtac aaatagtcta cataggtgtc tgggataatg 2880

taaaactggt gctttggctt tgtaaagaat ttgcaaatca cttaacagct gcaggggcaa 2940

ggggagagtt tcatcatccc catgatattt gggaatattc tgtttacttc ttagatagtt 3000

aagaatgtat tcagctacta tgtactaact tgaaccgtgt ttaaggaaaa ctcctatttc 3060

atcctcttct tgcgccatcc cctctcccta acttggtaat gtgaagaaac taaaacctga 3120

taccacagct cctataggca ttttagagat cttggatttt tatgtacagt cttagtcatt 3180

tttaataaat gtggttcagt aagggaa 3207

<210> 56

<211> 686

<212> PRT

<213> Intelligent

<400> 56

Met Gln Leu Gly Glu Gln Leu Leu Val Ser Ser Val Asn Leu Pro Gly

1 5 10 15

Ala His Phe Tyr Pro Leu Glu Ser Ala Arg Gly Gly Ser Gly Gly Ser

20 25 30

Ala Gly His Leu Pro Ser Ala Ala Pro Ser Pro Gln Lys Leu Asp Leu

35 40 45

Asp Lys Ala Ser Lys Lys Phe Ser Gly Ser Leu Ser Cys Glu Ala Val

50 55 60

Ser Gly Glu Pro Ala Ala Ala Ser Ala Gly Ala Pro Ala Ala Met Leu

65 70 75 80

Ser Asp Thr Asp Ala Gly Asp Ala Phe Ala Ser Ala Ala Ala Val Ala

85 90 95

Lys Pro Gly Pro Pro Asp Gly Arg Lys Gly Ser Pro Cys Gly Glu Glu

100 105 110

Glu Leu Pro Ser Ala Ala Ala Ala Ala Ala Ala Ala Ala Ala Ala Ala

115 120 125

Ala Ala Thr Ala Arg Tyr Ser Met Asp Ser Leu Ser Ser Glu Arg Tyr

130 135 140

Tyr Leu Gln Ser Pro Gly Pro Gln Gly Ser Glu Leu Ala Ala Pro Cys

145 150 155 160

Ser Leu Phe Pro Tyr Gln Ala Ala Ala Gly Ala Pro His Gly Pro Val

165 170 175

Tyr Pro Ala Pro Asn Gly Ala Arg Tyr Pro Tyr Gly Ser Met Leu Pro

180 185 190

Pro Gly Gly Phe Pro Ala Ala Val Cys Pro Pro Gly Arg Ala Gln Phe

195 200 205

Gly Pro Gly Ala Gly Ala Gly Ser Gly Ala Gly Gly Ser Ser Gly Gly

210 215 220

Gly Gly Gly Pro Gly Thr Tyr Gln Tyr Ser Gln Gly Ala Pro Leu Tyr

225 230 235 240

Gly Pro Tyr Pro Gly Ala Ala Ala Ala Gly Ser Cys Gly Gly Leu Gly

245 250 255

Gly Leu Gly Val Pro Gly Ser Gly Phe Arg Ala His Val Tyr Leu Cys

260 265 270

Asn Arg Pro Leu Trp Leu Lys Phe His Arg His Gln Thr Glu Met Ile

275 280 285

Ile Thr Lys Gln Gly Arg Arg Met Phe Pro Phe Leu Ser Phe Asn Ile

290 295 300

Asn Gly Leu Asn Pro Thr Ala His Tyr Asn Val Phe Val Glu Val Val

305 310 315 320

Leu Ala Asp Pro Asn His Trp Arg Phe Gln Gly Gly Lys Trp Val Thr

325 330 335

Cys Gly Lys Ala Asp Asn Asn Met Gln Gly Asn Lys Met Tyr Val His

340 345 350

Pro Glu Ser Pro Asn Thr Gly Ser His Trp Met Arg Gln Glu Ile Ser

355 360 365

Phe Gly Lys Leu Lys Leu Thr Asn Asn Lys Gly Ala Asn Asn Asn Asn

370 375 380

Thr Gln Met Ile Val Leu Gln Ser Leu His Lys Tyr Gln Pro Arg Leu

385 390 395 400

His Ile Val Glu Val Thr Glu Asp Gly Val Glu Asp Leu Asn Glu Pro

405 410 415

Ser Lys Thr Gln Thr Phe Thr Phe Ser Glu Thr Gln Phe Ile Ala Val

420 425 430

Thr Ala Tyr Gln Asn Thr Asp Ile Thr Gln Leu Lys Ile Asp His Asn

435 440 445

Pro Phe Ala Lys Gly Phe Arg Asp Asn Tyr Asp Ser Ser His Gln Ile

450 455 460

Val Pro Gly Gly Arg Tyr Gly Val Gln Ser Phe Phe Pro Glu Pro Phe

465 470 475 480

Val Asn Thr Leu Pro Gln Ala Arg Tyr Tyr Asn Gly Glu Arg Thr Val

485 490 495

Pro Gln Thr Asn Gly Leu Leu Ser Pro Gln Gln Ser Glu Glu Val Ala

500 505 510

Asn Pro Pro Gln Arg Trp Leu Val Thr Pro Val Gln Gln Pro Gly Thr

515 520 525

Asn Lys Leu Asp Ile Ser Ser Tyr Glu Ser Glu Tyr Thr Ser Ser Thr

530 535 540

Leu Leu Pro Tyr Gly Ile Lys Ser Leu Pro Leu Gln Thr Ser His Ala

545 550 555 560

Leu Gly Tyr Tyr Pro Asp Pro Thr Phe Pro Ala Met Ala Gly Trp Gly

565 570 575

Gly Arg Gly Ser Tyr Gln Arg Lys Met Ala Ala Gly Leu Pro Trp Thr

580 585 590

Ser Arg Thr Ser Pro Thr Val Phe Ser Glu Asp Gln Leu Ser Lys Glu

595 600 605

Lys Val Lys Glu Glu Ile Gly Ser Ser Trp Ile Glu Thr Pro Pro Ser

610 615 620

Ile Lys Ser Leu Asp Ser Asn Asp Ser Gly Val Tyr Thr Ser Ala Cys

625 630 635 640

Lys Arg Arg Arg Leu Ser Pro Ser Asn Ser Ser Asn Glu Asn Ser Pro

645 650 655

Ser Ile Lys Cys Glu Asp Ile Asn Ala Glu Glu Tyr Ser Lys Asp Thr

660 665 670

Ser Lys Gly Met Gly Gly Tyr Tyr Ala Phe Tyr Thr Thr Pro

675 680 685

<210> 57

<211> 884

<212> DNA

<213> Intelligent

<400> 57

cagccacaat gaggaactcc tatagatttc tggcatcctc tctctcagtt gtcgtttctc 60

tcctgctaat tcctgaagat gtctgtgaaa aaattattgg aggaaatgaa gtaactcctc 120

attcaagacc ctacatggtc ctacttagtc ttgacagaaa aaccatctgt gctggggctt 180

tgattgcaaa agactgggtg ttgactgcag ctcactgtaa cttgaacaaa aggtcccagg 240

tcattcttgg ggctcactca ataaccaggg aagagccaac aaaacagata atgcttgtta 300

agaaagagtt tccctatcca tgctatgacc cagccacacg cgaaggtgac cttaaacttt 360

tacagctgac ggaaaaagca aaaattaaca aatatgtgac tatccttcat ctacctaaaa 420

agggggatga tgtgaaacca ggaaccatgt gccaagttgc agggtggggc aggactcaca 480

atagtgcatc ttggtccgat actctgagag aagtcaatat caccatcata gacagaaaag 540

tctgcaatga tcgaaatcac tataatttta accctgtgat tggaatgaat atggtttgtg 600

ctggaagcct ccgaggtgga agagactcgt gcaatggaga ttctggaagc cctttgttgt 660

gcgagggtgt tttccgaggg gtcacttcct ttggccttga aaataaatgc ggagaccctc 720

gtgggcctgg tgtctatatt cttctctcaa agaaacacct caactggata attatgacta 780

tcaagggagc agtttaaata accgtttcct ttcatttact gtggcttctt aatcttttca 840

caaataaaat caatttgcat gactgtaaaa aaaaaaaaaa aaaa 884

<210> 58

<211> 262

<212> PRT

<213> Intelligent

<400> 58

Met Arg Asn Ser Tyr Arg Phe Leu Ala Ser Ser Leu Ser Val Val Val

1 5 10 15

Ser Leu Leu Leu Ile Pro Glu Asp Val Cys Glu Lys Ile Ile Gly Gly

20 25 30

Asn Glu Val Thr Pro His Ser Arg Pro Tyr Met Val Leu Leu Ser Leu

35 40 45

Asp Arg Lys Thr Ile Cys Ala Gly Ala Leu Ile Ala Lys Asp Trp Val

50 55 60

Leu Thr Ala Ala His Cys Asn Leu Asn Lys Arg Ser Gln Val Ile Leu

65 70 75 80

Gly Ala His Ser Ile Thr Arg Glu Glu Pro Thr Lys Gln Ile Met Leu

85 90 95

Val Lys Lys Glu Phe Pro Tyr Pro Cys Tyr Asp Pro Ala Thr Arg Glu

100 105 110

Gly Asp Leu Lys Leu Leu Gln Leu Met Glu Lys Ala Lys Ile Asn Lys

115 120 125

Tyr Val Thr Ile Leu His Leu Pro Lys Lys Gly Asp Asp Val Lys Pro

130 135 140

Gly Thr Met Cys Gln Val Ala Gly Trp Gly Arg Thr His Asn Ser Ala

145 150 155 160

Ser Trp Ser Asp Thr Leu Arg Glu Val Asn Ile Thr Ile Ile Asp Arg

165 170 175

Lys Val Cys Asn Asp Arg Asn His Tyr Asn Phe Asn Pro Val Ile Gly

180 185 190

Met Asn Met Val Cys Ala Gly Ser Leu Arg Gly Gly Arg Asp Ser Cys

195 200 205

Asn Gly Asp Ser Gly Ser Pro Leu Leu Cys Glu Gly Val Phe Arg Gly

210 215 220

Val Thr Ser Phe Gly Leu Glu Asn Lys Cys Gly Asp Pro Arg Gly Pro

225 230 235 240

Gly Val Tyr Ile Leu Leu Ser Lys Lys His Leu Asn Trp Ile Ile Met

245 250 255

Thr Ile Lys Gly Ala Val

260

<210> 59

<211> 2583

<212> DNA

<213> Intelligent people

<400> 59

agtgacagcg gcccgctgga gaggaagccc gagagctgcc gcgcgcctgc cggacgaggg 60

cgtagaagcc aggcgtcaga gcccgggctc cggtggggtc ccccacccgg ccctcgggtc 120

ccccgccccc tgctccctgc ccatcccagc ccacgcgacc ctctcgcgcg cggaggggcg 180

ggtcctcgac ggctacggga aggtgccagc ccgccccgga tgggcatcgt ggagccgggt 240

tgcggagaca tgctgacggg caccgagccg atgccgggga gcgacgaggg ccgggcgcct 300

ggcgccgacc cgcagcaccg ctacttctac ccggagccgg gcgcgcagga cgcggacgag 360

cgtcgcgggg gcggcagcct ggggtctccc tacccggggg gcgccttggt gcccgccccg 420

ccgagccgct tccttggagc ctacgcctac ccgccgcgac cccaggcggc cggcttcccc 480

ggcgcgggcg agtccttccc gccgcccgcg gacgccgagg gctaccagcc gggcgagggc 540

tacgccgccc cggacccgcg cgccgggctc tacccggggc cgcgtgagga ctacgcgcta 600

cccgcgggac tggaggtgtc ggggaaactg agggtcgcgc tcaacaacca cctgttgtgg 660

tccaagttta atcagcacca gacagagatg atcatcacca agcagggacg gcggatgttc 720

ccattcctgt catttactgt ggccgggctg gagcccacca gccactacag gatgtttgtg 780

gacgtggtct tggtggacca gcaccactgg cggtaccaga gcggcaagtg ggtgcagtgt 840

ggaaaggccg agggcagcat gccaggaaac cgcctgtacg tccacccgga ctcccccaac 900

acaggagcgc actggatgcg ccaggaagtt tcatttggga aactaaagct cacaaacaac 960

aagggggcgt ccaacaatgt gacccagatg attgtgctcc agtccctcca taagtaccag 1020

ccccggctgc atatcgttga ggtgaacgac ggagagccag aggcagcctg caacgcttcc 1080

aacacgcata tctttacttt ccaagaaacc cagttcattg ccgtgactgc ctaccagaat 1140

gccgagatta ctcagctgaa aattgataat aacccctttg ccaaaggatt ccgggagaac 1200

tttgagtcca tgtacacatc tgttgacacc agcatcccct ccccgcctgg acccaactgt 1260

caattccttg ggggagatca ctactctcct ctcctaccca accagtatcc tgttcccagc 1320

cgcttctacc ccgaccttcc tggccaggcg aaggatgtgg ttccccaggc ttactggctg 1380

ggggcccccc gggaccacag ctatgaggct gagtttcgag cagtcagcat gaagcctgca 1440

ttcttgccct ctgcccctgg gcccaccatg tcctactacc gaggccagga ggtcctggca 1500

cctggagctg gctggcctgt ggcaccccag taccctccca agatgggccc ggccagctgg 1560

ttccgcccta tgcggactct gcccatggaa cccggccctg gaggctcaga gggacgggga 1620

ccagaggacc agggtccccc cttggtgtgg actgagattg cccccatccg gccggaatcc 1680

agtgattcag gactgggcga aggagactct aagaggaggc gcgtgtcccc ctatccttcc 1740

agtggtgaca gctcctcccc tgctggggcc ccttctcctt ttgataagga agctgaagga 1800

cagttttata actattttcc caactgagca gatgacatga tgaaaggaac agaaacagtg 1860

ttattaggtt ggaggacacc gactaatttg ggaaacggat gaaggactga gaaggccccc 1920

gctccctctg gcccttctct gtttagtagt tggttgggga agtggggctc aagaaggatt 1980

ttggggttca ccagatgctt cctggcccac gatgaaacct gagaggggtg tccccttgcc 2040

ccatcctctg ccctaactac agtcgtttac ctggtgctgc gtcttgcttt tggtttccag 2100

ctggagaaaa gaagacaaga aagtcttggg catgaaggag ctttttgcat ctagtgggtg 2160

ggaggggtca ggtgtgggac atgggagcag gagactccac tttcttcctt tgtacagtaa 2220

ctttcaacct tttcgttggc atgtgtgtta atccctgatc caaaaagaac aaatacacgt 2280

atgttataac catcagcccg ccagggtcag ggaaaggact cacctgactt tggacagctg 2340

gcctgggctc cccctgctca aacacagtgg ggatcagaga aaaggggctg gaaagggggg 2400

aatggcccac atctcaagaa gcaagatatt gtttgtggtg gttgtgtgtg ggtgtgtgtt 2460

ttttcttttt ctttcttttt attttttttg aatgggggag gctatttatt gtactgagag 2520

tggtgtctgg atatattcct tttgtcttca tcactttctg aaaataaaca taaaactgtt 2580

gaa 2583

<210> 60

<211> 535

<212> PRT

<213> Intelligent

<400> 60

Met Gly Ile Val Glu Pro Gly Cys Gly Asp Met Leu Thr Gly Thr Glu

1 5 10 15

Pro Met Pro Gly Ser Asp Glu Gly Arg Ala Pro Gly Ala Asp Pro Gln

20 25 30

His Arg Tyr Phe Tyr Pro Glu Pro Gly Ala Gln Asp Ala Asp Glu Arg

35 40 45

Arg Gly Gly Gly Ser Leu Gly Ser Pro Tyr Pro Gly Gly Ala Leu Val

50 55 60

Pro Ala Pro Pro Ser Arg Phe Leu Gly Ala Tyr Ala Tyr Pro Pro Arg

65 70 75 80

Pro Gln Ala Ala Gly Phe Pro Gly Ala Gly Glu Ser Phe Pro Pro Pro

85 90 95

Ala Asp Ala Glu Gly Tyr Gln Pro Gly Glu Gly Tyr Ala Ala Pro Asp

100 105 110

Pro Arg Ala Gly Leu Tyr Pro Gly Pro Arg Glu Asp Tyr Ala Leu Pro

115 120 125

Ala Gly Leu Glu Val Ser Gly Lys Leu Arg Val Ala Leu Asn Asn His

130 135 140

Leu Leu Trp Ser Lys Phe Asn Gln His Gln Thr Glu Met Ile Ile Thr

145 150 155 160

Lys Gln Gly Arg Arg Met Phe Pro Phe Leu Ser Phe Thr Val Ala Gly

165 170 175

Leu Glu Pro Thr Ser His Tyr Arg Met Phe Val Asp Val Val Leu Val

180 185 190

Asp Gln His His Trp Arg Tyr Gln Ser Gly Lys Trp Val Gln Cys Gly

195 200 205

Lys Ala Glu Gly Ser Met Pro Gly Asn Arg Leu Tyr Val His Pro Asp

210 215 220

Ser Pro Asn Thr Gly Ala His Trp Met Arg Gln Glu Val Ser Phe Gly

225 230 235 240

Lys Leu Lys Leu Thr Asn Asn Lys Gly Ala Ser Asn Asn Val Thr Gln

245 250 255

Met Ile Val Leu Gln Ser Leu His Lys Tyr Gln Pro Arg Leu His Ile

260 265 270

Val Glu Val Asn Asp Gly Glu Pro Glu Ala Ala Cys Asn Ala Ser Asn

275 280 285

Thr His Ile Phe Thr Phe Gln Glu Thr Gln Phe Ile Ala Val Thr Ala

290 295 300

Tyr Gln Asn Ala Glu Ile Thr Gln Leu Lys Ile Asp Asn Asn Pro Phe

305 310 315 320

Ala Lys Gly Phe Arg Glu Asn Phe Glu Ser Met Tyr Thr Ser Val Asp

325 330 335

Thr Ser Ile Pro Ser Pro Pro Gly Pro Asn Cys Gln Phe Leu Gly Gly

340 345 350

Asp His Tyr Ser Pro Leu Leu Pro Asn Gln Tyr Pro Val Pro Ser Arg

355 360 365

Phe Tyr Pro Asp Leu Pro Gly Gln Ala Lys Asp Val Val Pro Gln Ala

370 375 380

Tyr Trp Leu Gly Ala Pro Arg Asp His Ser Tyr Glu Ala Glu Phe Arg

385 390 395 400

Ala Val Ser Met Lys Pro Ala Phe Leu Pro Ser Ala Pro Gly Pro Thr

405 410 415

Met Ser Tyr Tyr Arg Gly Gln Glu Val Leu Ala Pro Gly Ala Gly Trp

420 425 430

Pro Val Ala Pro Gln Tyr Pro Pro Lys Met Gly Pro Ala Ser Trp Phe

435 440 445

Arg Pro Met Arg Thr Leu Pro Met Glu Pro Gly Pro Gly Gly Ser Glu

450 455 460

Gly Arg Gly Pro Glu Asp Gln Gly Pro Pro Leu Val Trp Thr Glu Ile

465 470 475 480

Ala Pro Ile Arg Pro Glu Ser Ser Asp Ser Gly Leu Gly Glu Gly Asp

485 490 495

Ser Lys Arg Arg Arg Val Ser Pro Tyr Pro Ser Ser Gly Asp Ser Ser

500 505 510

Ser Pro Ala Gly Ala Pro Ser Pro Phe Asp Lys Glu Ala Glu Gly Gln

515 520 525

Phe Tyr Asn Tyr Phe Pro Asn

530 535

<210> 61

<211> 1211

<212> DNA

<213> Intelligent

<400> 61

acattgttct gatcatctga agatcagcta ttagaagaga aagatcagtt aagtcctttg 60

gacctgatca gcttgataca agaactactg atttcaactt ctttggctta attctctcgg 120

aaacgatgaa atatacaagt tatatcttgg cttttcagct ctgcatcgtt ttgggttctc 180

ttggctgtta ctgccaggac ccatatgtaa aagaagcaga aaaccttaag aaatatttta 240

atgcaggtca ttcagatgta gcggataatg gaactctttt cttaggcatt ttgaagaatt 300

ggaaagagga gagtgacaga aaaataatgc agagccaaat tgtctccttt tacttcaaac 360

tttttaaaaa ctttaaagat gaccagagca tccaaaagag tgtggagacc atcaaggaag 420

acatgaatgt caagtttttc aatagcaaca aaaagaaacg agatgacttc gaaaagctga 480

ctaattattc ggtaactgac ttgaatgtcc aacgcaaagc aatacatgaa ctcatccaag 540

tgatggctga actgtcgcca gcagctaaaa cagggaagcg aaaaaggagt cagatgctgt 600

ttcgaggtcg aagagcatcc cagtaatggt tgtcctgcct gcaatatttg aattttaaat 660

ctaaatctat ttattaatat ttaacattat ttatatgggg aatatatttt tagactcatc 720

aatcaaataa gtatttataa tagcaacttt tgtgtaatga aaatgaatat ctattaatat 780

atgtattatt tataattcct atatcctgtg actgtctcac ttaatccttt gttttctgac 840

taattaggca aggctatgtg attacaaggc tttatctcag gggccaacta ggcagccaac 900

ctaagcaaga tcccatgggt tgtgtgttta tttcacttga tgatacaatg aacacttata 960

agtgaagtga tactatccag ttactgccgg tttgaaaata tgcctgcaat ctgagccagt 1020

gctttaatgg catgtcagac agaacttgaa tgtgtcaggt gaccctgatg aaaacatagc 1080

atctcaggag atttcatgcc tggtgcttcc aaatattgtt gacaactgtg actgtaccca 1140

aatggaaagt aactcatttg ttaaaattat caatatctaa tatatatgaa taaagtgtaa 1200

gttcacaact a 1211

<210> 62

<211> 166

<212> PRT

<213> Intelligent

<400> 62

Met Lys Tyr Thr Ser Tyr Ile Leu Ala Phe Gln Leu Cys Ile Val Leu

1 5 10 15

Gly Ser Leu Gly Cys Tyr Cys Gln Asp Pro Tyr Val Lys Glu Ala Glu

20 25 30

Asn Leu Lys Lys Tyr Phe Asn Ala Gly His Ser Asp Val Ala Asp Asn

35 40 45

Gly Thr Leu Phe Leu Gly Ile Leu Lys Asn Trp Lys Glu Glu Ser Asp

50 55 60

Arg Lys Ile Met Gln Ser Gln Ile Val Ser Phe Tyr Phe Lys Leu Phe

65 70 75 80

Lys Asn Phe Lys Asp Asp Gln Ser Ile Gln Lys Ser Val Glu Thr Ile

85 90 95

Lys Glu Asp Met Asn Val Lys Phe Phe Asn Ser Asn Lys Lys Lys Arg

100 105 110

Asp Asp Phe Glu Lys Leu Thr Asn Tyr Ser Val Thr Asp Leu Asn Val

115 120 125

Gln Arg Lys Ala Ile His Glu Leu Ile Gln Val Met Ala Glu Leu Ser

130 135 140

Pro Ala Ala Lys Thr Gly Lys Arg Lys Arg Ser Gln Met Leu Phe Arg

145 150 155 160

Gly Arg Arg Ala Ser Gln

165

<210> 63

<211> 884

<212> DNA

<213> Intelligent

<400> 63

cagccacaat gaggaactcc tatagatttc tggcatcctc tctctcagtt gtcgtttctc 60

tcctgctaat tcctgaagat gtctgtgaaa aaattattgg aggaaatgaa gtaactcctc 120

attcaagacc ctacatggtc ctacttagtc ttgacagaaa aaccatctgt gctggggctt 180

tgattgcaaa agactgggtg ttgactgcag ctcactgtaa cttgaacaaa aggtcccagg 240

tcattcttgg ggctcactca ataaccaggg aagagccaac aaaacagata atgcttgtta 300

agaaagagtt tccctatcca tgctatgacc cagccacacg cgaaggtgac cttaaacttt 360

tacagctgac ggaaaaagca aaaattaaca aatatgtgac tatccttcat ctacctaaaa 420

agggggatga tgtgaaacca ggaaccatgt gccaagttgc agggtggggc aggactcaca 480

atagtgcatc ttggtccgat actctgagag aagtcaatat caccatcata gacagaaaag 540

tctgcaatga tcgaaatcac tataatttta accctgtgat tggaatgaat atggtttgtg 600

ctggaagcct ccgaggtgga agagactcgt gcaatggaga ttctggaagc cctttgttgt 660

gcgagggtgt tttccgaggg gtcacttcct ttggccttga aaataaatgc ggagaccctc 720

gtgggcctgg tgtctatatt cttctctcaa agaaacacct caactggata attatgacta 780

tcaagggagc agtttaaata accgtttcct ttcatttact gtggcttctt aatcttttca 840

caaataaaat caatttgcat gactgtaaaa aaaaaaaaaa aaaa 884

<210> 64

<211> 247

<212> PRT

<213> Intelligent

<400> 64

Met Gln Pro Ile Leu Leu Leu Leu Ala Phe Leu Leu Leu Pro Arg Ala

1 5 10 15

Asp Ala Gly Glu Ile Ile Gly Gly His Glu Ala Lys Pro His Ser Arg

20 25 30

Pro Tyr Met Ala Tyr Leu Met Ile Trp Asp Gln Lys Ser Leu Lys Arg

35 40 45

Cys Gly Gly Phe Leu Ile Arg Asp Asp Phe Val Leu Thr Ala Ala His

50 55 60

Cys Trp Gly Ser Ser Ile Asn Val Thr Leu Gly Ala His Asn Ile Lys

65 70 75 80

Glu Gln Glu Pro Thr Gln Gln Phe Ile Pro Val Lys Arg Pro Ile Pro

85 90 95

His Pro Ala Tyr Asn Pro Lys Asn Phe Ser Asn Asp Ile Met Leu Leu

100 105 110

Gln Leu Glu Arg Lys Ala Lys Arg Thr Arg Ala Val Gln Pro Leu Arg

115 120 125

Leu Pro Ser Asn Lys Ala Gln Val Lys Pro Gly Gln Thr Cys Ser Val

130 135 140

Ala Gly Trp Gly Gln Thr Ala Pro Leu Gly Lys His Ser His Thr Leu

145 150 155 160

Gln Glu Val Lys Met Thr Val Gln Glu Asp Arg Lys Cys Glu Ser Asp

165 170 175

Leu Arg His Tyr Tyr Asp Ser Thr Ile Glu Leu Cys Val Gly Asp Pro

180 185 190

Glu Ile Lys Lys Thr Ser Phe Lys Gly Asp Ser Gly Gly Pro Leu Val

195 200 205

Cys Asn Lys Val Ala Gln Gly Ile Val Ser Tyr Gly Arg Asn Asn Gly

210 215 220

Met Pro Pro Arg Ala Cys Thr Lys Val Ser Ser Phe Val His Trp Ile

225 230 235 240

Lys Lys Thr Met Lys Arg Tyr

245

<210> 65

<211> 3394

<212> DNA

<213> Intelligent

<400> 65

gtgcttgaga aggttcaatg gcgtggcagg gactagcggc cgagttcctg caggtgccgg 60

cggtgacgcg ggcttacacc gcagcctgtg tcctcaccac cgccgcggtg gtaagcggcc 120

gggcggaccg gacgtcgcct tggttacagc ctctggcggg aggggtgagg gtcgccatgg 180

ttacggcgtg gctcccgggc agctcttggc tggcccctgg ttccacagca gctggagctc 240

ctcagcccct ttcaactcta cttcaacccg caccttgtgt tccggaagtt ccaggtctgg 300

aggctcgtca ccaacttcct cttcttcggg cccctgggat tcagcttctt cttcaacatg 360

ctcttcgtgt tccgctactg ccgcatgctg gaagagggct ccttccgcgg ccgcacggcc 420

gacttcgtct tcatgtttct cttcgggggc gtccttatga ccgtatcctt cccgcaggct 480

ctggaacctc gggctagggc gcctcggcgt ccagcctgtg ttggtcctgg ggccaacaca 540

gccatgccag aaagggacac agtcgctgtc tccagcttag tatgtgttga gggcccactc 600

tgtgctcagc tgcaggggtc agggctagat cttcagtgct gtatgcaaaa tacaaagcca 660

cgcacaaaag agccaggcac cgttcctgcc ttgggcgctc atgggcttct cgctgctgct 720

gggcaactcc atcctcgtgg acctgctggg gattgcggtg ggccatatct actacttcct 780

ggaggacgtc ttccccaacc agcctggagg caagaggctc ctgcagaccc ctggcttcct 840

gtgagtgttg agagccctcc ctccctctcc ccaccctcag aaggatcccc accgatgggg 900

acctgtgctg gcctgtgctc aacacgggcc cctccccaca gaaagctgct cctggatgtc 960

cctgcagaag accccaatta cctgcccctc cctgaggaac agccaggacc ccatctgcca 1020

cccccgcagc agtgaccccc acccagggcc aggcctaaga ggcttctggc agcttccatc 1080

ctacccatga cccctacttg gggcagaaaa aacccatcct aaaggctggg cccatgcaag 1140

ggcccacctg aataaacaga atgagctgca gtctcttggc ccacagcact ggcttcccca 1200

tctcacctgg ccacatcctt ctatgcctgc cccgtcctca ctcagtgtgg cctctcagcc 1260

caactgcagg tggtaggata ggggtgccca cagagggcaa agaaactgcc catggttgcc 1320

tggcagagct ttgagctcac aggttgccag gcagagcttt tgagctcaca ggtgacaggc 1380

tcagggttct catcctggcc ccaccagggc cttgggcaag tcctgcccac cataggcctc 1440

tgctacctgc cagccagcgg ggaagttcac cagatttcgg ctgctggggc caggacaggc 1500

ctctcctagg ttgtgccaaa ccagcctaca gatgttcctg ccagtggtgc cttcaggctg 1560

atgccaatct agcctctcct gtctctcatc agccaccctg acaggtgggc gtatgcctca 1620

tttttcatct ggtgatgcca aagccccatg gattcaggtg cagaagaggg ccaggactag 1680

gtcttctgcc ctttctatga cctcagagcc taagttttca ccttagcaga gttctgagac 1740

tgggtgaggc agggacttct ggaaggttct gttcctgccc tttttagctg aggacgtgtg 1800

tgagccttat ccgacccctg tggctcattt ttctcttctg acctggcagc tttccttgtt 1860

gttctaagcc tgtccatgtt gtggtttatt tctggatgct cagtggcacg gggcctcctc 1920

caaagacagg ttgtcatttt catggtaaca acactgttct ctgttgagtc tgccctccgt 1980

gttgtagcca gaccttgtgg agatggcttt gcggcggtgt gagctggcgg tcaggagtac 2040

ccagccttcc cggcacctcc cagccaggtg gccctgcccg acctgtgggg tgaggcagcc 2100

aaggttcctt ccatcccccc agttgtggag acacagggct gcctcaccct ttcattgccg 2160

aggttcctac ctcatggaca gaacaaacac ctcagcaatg aaacctgttc atgtctaaga 2220

gcagctgggc tgggaatctt cccctttgtt caaggccttc cagtaaggcc cagctgtccc 2280

cttgctgtgc atggggctct ggggagttcc actctttgat ggagggcaga ggccctgagt 2340

gcaaactccc tgggaagagt cccatgctaa catgtgctca aaggagcccc cctctcacat 2400

ctcagcgacc aagaacccca atccctaatc agagctctgc ctctgcccca tatggggccc 2460

taccccactt ccagggagca cagcagcctc tgaccctagc cctgccctgg cagcatggga 2520

cctgccaaca gctgagggtg gcagcagtct gtgctgggat ctgtgcccgg atctgtgccc 2580

attctcttca gtaaggctgg agctgcgagc cagtttgctg ccctcccaga tcctgtgtcc 2640

attctgaaga atggggacac ctccctatta cagatgagaa cagaggggac atgaactcct 2700

tggaggcagg gctgggaagg gaccctgggc tgtgtctcct ccctgtaccg tgtcaactcc 2760

aagagctggc accaggccgg gagggctatt cccatattcc tcacagctgg cttgtaaggc 2820

aggagctggc ccaggccaca gcacctgctg gggtgggaaa gggccccagc tgacctggct 2880

atgggtgcca ctggagctag aacagccctc ctgcacccag gctggcgaca gcccagcagt 2940

gcccacacag gactgggccc tccgcagggg acttcagagc agcaaggccc cagctggcag 3000

tagcctgacc atctggacac agcagagcca gggcggccca gggacggcag gagagctcgc 3060

ggcaccttcc tgaggccaag caaggggagc agggttaggg ctgttcctga aaggcagaga 3120

gcctctgccc tgagcctcac agctactctt ctcagctctc tgggtctgga aggagaacag 3180

gctgagggga gctgagagga gctgaggtgc tacccggagc cccattcacc cccacctgcc 3240

cacttgggaa tctgaggcag aggagggtga ggcctgtgtg ccaaccttgt tcacatacca 3300

ccttcgtccc cccaggcccc ggccccactc ctggctctca ttatttttat gttaaaactt 3360

tgaagaaatt gaacatgacc tgttgaagaa attg 3394

<210> 66

<211> 235

<212> PRT

<213> Intelligent

<400> 66

Met Ala Trp Gln Gly Leu Ala Ala Glu Phe Leu Gln Val Pro Ala Val

1 5 10 15

Thr Arg Ala Tyr Thr Ala Ala Cys Val Leu Thr Thr Ala Ala Val Gln

20 25 30

Leu Glu Leu Leu Ser Pro Phe Gln Leu Tyr Phe Asn Pro His Leu Val

35 40 45

Phe Arg Lys Phe Gln Val Trp Arg Leu Val Thr Asn Phe Leu Phe Phe

50 55 60

Gly Pro Leu Gly Phe Ser Phe Phe Phe Asn Met Leu Phe Val Phe Arg

65 70 75 80

Tyr Cys Arg Met Leu Glu Glu Gly Ser Phe Arg Gly Arg Thr Ala Asp

85 90 95

Phe Val Phe Met Phe Leu Phe Gly Gly Val Leu Met Thr Leu Leu Gly

100 105 110

Leu Leu Gly Ser Leu Phe Phe Leu Gly Gln Ala Leu Met Ala Met Leu

115 120 125

Val Tyr Val Trp Ser Arg Arg Ser Pro Arg Val Arg Val Asn Phe Phe

130 135 140

Gly Leu Leu Thr Phe Gln Ala Pro Phe Leu Pro Trp Ala Leu Met Gly

145 150 155 160

Phe Ser Leu Leu Leu Gly Asn Ser Ile Leu Val Asp Leu Leu Gly Ile

165 170 175

Ala Val Gly His Ile Tyr Tyr Phe Leu Glu Asp Val Phe Pro Asn Gln

180 185 190

Pro Gly Gly Lys Arg Leu Leu Gln Thr Pro Gly Phe Leu Lys Leu Leu

195 200 205

Leu Asp Ala Pro Ala Glu Asp Pro Asn Tyr Leu Pro Leu Pro Glu Glu

210 215 220

Gln Pro Gly Pro His Leu Pro Pro Pro Gln Gln

225 230 235

<210> 67

<211> 1173

<212> DNA

<213> Intelligent

<400> 67

ggcggcctgg cccgggacct gctgctgctc cagccatgtc catgacaacc agagcctggg 60

aggagctgga tggcggcctg ggcagctgcc aggccctgga ggaccactct gcgctggccg 120

agacccagga ggacagggct tcagcgacac ccaggctggc cgactccggc agcgtgcccc 180

acgactctca ggtggctgaa ggccccagtg tggacaccag gcccaagaag atggaaaaag 240

agcctgccgc cagggggacc ccaggaacgg ggaaggagag gctgaaagcc ggagcgagcc 300

ctcggagcgt ccccgcgcgc aagaaggcgc agaccgcgcc gcccctgcag ccgccgccgc 360

cgcccccggc cctgagcgag gagctgccct ggggagacct gtcgctcaac aagtgcctgg 420

tgctcgcctc gctggtggcg ctgctgggct cggctttcca gctgtgccgc gacgccgtcc 480

ctggggaggc agcactccaa gcacgtgtgc ccgagccctg ggtcccgcca agctcagccc 540

cgagggagcc atcgtcgccc ctgcctaagt tcgaggccca ggcgcctcca tcagcgccgc 600

ctgcgccccg ggccgaggca gaggtcagac ccaagattcc cgggagtcgg gaggctgcag 660

agaacgacga agaggagccc ggcgaggcca ccggagaggc cgtccgggag gaccgtgtga 720

ccctcgcaga ccggggaccc aaggagaggc ctcggagaga ggggaagccg cggaaggaga 780

agccgcggaa ggaggagaga cctaagaaag agaggccgcg gaaagaggag aggccacggg 840

ccgccaggga gccccgggaa gccctacccc agcgctggga gtcacgcgaa gggggccacc 900

ggccgtgggc acgggactcc agggacgccg agcccaggaa gaagcaggcc tgggtgtccc 960

cgaggcgtcc cgacgaggag cagcggcctg ggagtcgcca gaagctccgc gcaggcaagg 1020

ggcgggactg agccggcccc gcgccggagt ccaggggccc cttctggacg ccccgcgact 1080

ctggcgaaat aaagcgagtg ctgcggcaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 1140

aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaa 1173

<210> 68

<211> 331

<212> PRT

<213> Intelligent

<400> 68

Met Ser Met Thr Thr Arg Ala Trp Glu Glu Leu Asp Gly Gly Leu Gly

1 5 10 15

Ser Cys Gln Ala Leu Glu Asp His Ser Ala Leu Ala Glu Thr Gln Glu

20 25 30

Asp Arg Ala Ser Ala Thr Pro Arg Leu Ala Asp Ser Gly Ser Val Pro

35 40 45

His Asp Ser Gln Val Ala Glu Gly Pro Ser Val Asp Thr Arg Pro Lys

50 55 60

Lys Met Glu Lys Glu Pro Ala Ala Arg Gly Thr Pro Gly Thr Gly Lys

65 70 75 80

Glu Arg Leu Lys Ala Gly Ala Ser Pro Arg Ser Val Pro Ala Arg Lys

85 90 95

Lys Ala Gln Thr Ala Pro Pro Leu Gln Pro Pro Pro Pro Pro Pro Ala

100 105 110

Leu Ser Glu Glu Leu Pro Trp Gly Asp Leu Ser Leu Asn Lys Cys Leu

115 120 125

Val Leu Ala Ser Leu Val Ala Leu Leu Gly Ser Ala Phe Gln Leu Cys

130 135 140

Arg Asp Ala Val Pro Gly Glu Ala Ala Leu Gln Ala Arg Val Pro Glu

145 150 155 160

Pro Trp Val Pro Pro Ser Ser Ala Pro Arg Glu Pro Ser Ser Pro Leu

165 170 175

Pro Lys Phe Glu Ala Gln Ala Pro Pro Ser Ala Pro Pro Ala Pro Arg

180 185 190

Ala Glu Ala Glu Val Arg Pro Lys Ile Pro Gly Ser Arg Glu Ala Ala

195 200 205

Glu Asn Asp Glu Glu Glu Pro Gly Glu Ala Thr Gly Glu Ala Val Arg

210 215 220

Glu Asp Arg Val Thr Leu Ala Asp Arg Gly Pro Lys Glu Arg Pro Arg

225 230 235 240

Arg Glu Gly Lys Pro Arg Lys Glu Lys Pro Arg Lys Glu Glu Arg Pro

245 250 255

Lys Lys Glu Arg Pro Arg Lys Glu Glu Arg Pro Arg Ala Ala Arg Glu

260 265 270

Pro Arg Glu Ala Leu Pro Gln Arg Trp Glu Ser Arg Glu Gly Gly His

275 280 285

Arg Pro Trp Ala Arg Asp Ser Arg Asp Ala Glu Pro Arg Lys Lys Gln

290 295 300

Ala Trp Val Ser Pro Arg Arg Pro Asp Glu Glu Gln Arg Pro Gly Ser

305 310 315 320

Arg Gln Lys Leu Arg Ala Gly Lys Gly Arg Asp

325 330

<210> 69

<211> 32230

<212> DNA

<213> Intelligent

<400> 69

gaattcactc atcaccaagg ggaagatgct caatcattca tgagggatct gcccccttga 60

tctaatcacc tcctagtggc cccacttcca acactgggaa tcatattaca acatgagatt 120

tggaggggac aaatatataa accatatcct tccacccctg gtcccccaaa tctcatgtcc 180

ttctcatgtt gcaaaataca atcatgcctt ctcaacagtc ccccaaagtc ttacctcatt 240

ccagaatcaa caaaaaattc ccaagtccca agtctcatct gaagatgagt cccttccacc 300

tatcagccta ggaaatcaaa gacaaattga ctcccaaaat acaatgagag tgcagacatt 360

gggtaaacat tcccattcca caagggagaa gttggcaaaa agaaaggatc tacatgcccc 420

atacacgtca gaaatccagc agggcagtta ttaaatctta aagctccaag gtaatctcct 480

ttgacttcat gtcccatatc cagggcacac tggtgcaggg ggtgggctcc caaagccttg 540

gacaactcta cccctgtggc tttggaggat gtcgcccctg tggcttctct cacacgatag 600

agttgagtgc ctgtggcttt tccagattca gggtgcaaac tgccagtgga tctaccattc 660

tggggtctgg aggacagtgg cccccttccc acagctccac taggcagtgc cctggtgggg 720

acattatatg ggggttcaac cccacattct ctttggcact gctctagtag aggttctctg 780

tgagggctcc atgctggcag gaggcttctc tctgggcacc tagggtttct catacatttc 840

tgaaatctag agtgaagatg ccaagcctcc ttcactcttg cattctgggg gcctacaggc 900

ttaacaccag atggaagttc ccaaggctta tagtggcttg cactctccaa agaagcagcc 960

taagctgtac ttggggccct ttgagccaag gctggagcca gaggaaccag gatgtggaga 1020

gcagtgtccc aaggctgcac aggcagcagt ggccatgggc atggcccaca aaaccatcct 1080

ttccctcttg gcctctgggc ctgttatggg aaagaccacc tcagagattt ccgaaaggcc 1140

ttcaaggcct ttttcccttt gttctggata tgagcactta gctccctttt agttatgcta 1200

atctctctag caactggctg ctccatagcc tacttatatt tgtctcctga aaataccttt 1260

tcttttctac cacatggcca ggctgtgaat tttccacatt ttttatgctc tgcttccctt 1320

tgttgatgaa aaaagccaaa ctccataaaa cacttgacga gattgattct gagccacatg 1380

tgagagccat gaactgtggc acagcctcaa aaggtcctga gaacacctgt gcaaggtggt 1440

tgggttgcgg cctgctttta tgttttaggg agacatatca atcaatacat gtaaagtata 1500

cattggtttg gtttggtttg gaaaagcagg acaattcaaa gtggggaatt ccaaatcata 1560

ggtggatttg aagattttct gattggcaat tggctgaaaa agttaaatta tctaaaaagt 1620

tgaagtcagc aaaaagcaat gcttaagata agggggttgt ggaagccaag tttcttgtta 1680

tgtagatgaa gcctccacgt tccagagaga agagatggtc aatgtctctt atcagaacct 1740

aaaaggtgcc agactcttgg ataaatctct cctgaatcag gaagagacct ggaaagggaa 1800

aaagattctc aacagaatat acatttcctc cacaagagac ggctttgcag ggcccttcca 1860

aaatgtgtca gagaaatata ttctggcata aaatacttta atttccttcc atgcctgcca 1920

cctgtcatgt gatgctatac cagaatcagg ttgggatttg atatcttatt gctacaaaga 1980

gtctgctttg tcagtcttaa gctccctgtt ttaatgttaa cactggtcag ctgagcctaa 2040

gctccaacag ggagagggta tagtgaggga gtccaaacca ccctcccctt cctgtcatgg 2100

cctgaactgg ttttccaggt ttctttggaa tccccttgga ttagcatttt attttgtcgt 2160

ttacaccttt taaatataag ttccaacttt aagtcatttc tttgctccca tatcttacca 2220

tagactgtta gaagcaacca ggtcacacct tagaaacatc tgcttagaaa tgtcttccac 2280

agatgcccta agtcatcact cttaaggtca accttccaca gatccctagg gcatgaacac 2340

aatgcagcca agttgtttct taggatttaa caagcgtgac ctttacacca gttcccaata 2400

agtttctcat ttccatctga gatttcatca gcctggcttt cacagtccat atgtctatca 2460

gcattttggt gacaaccatt taatagtctc taagaaattc caaactttcc ctcatcttcc 2520

tgtcttcttc tgagtcctcc aaattcttct aacctctctg cctgttacct gattccaaag 2580

ctgcttccac attttcaggt atccttatag caatgttcca ctcctcaata ccaatgttct 2640

gagttagtct atttgtgtta ctagaaaaaa aaaaaaaaca tgaatctggt aatttataaa 2700

gaaaagaggt ttgcaccagg cactgtggca aggaagggag gattaactag ctcacagttc 2760

tgcagccatt ccggaaacac ggccctggca tctgcttctg gcgaggcctc agggagattg 2820

caatcacggc agaaggcgaa gggggagcag gcatgtcaca tggcaagagg gagcaagaga 2880

agagaagtgg cacgtcctag acttttaaac aaccagctgt cacagggact aactgaggga 2940

gaactcactc atcacctggg agatggtgct aggccattca tgaggggtcc agccccatga 3000

tctaatcacc tcccaccagg cccacctcca acactgggaa tcacatctca acatgaaatt 3060

tggagagaca cacatccaaa ctattgcacc acaggaagct ggaagaggtg gacggtcctc 3120

ccctggagca ctcagagaca gccaccaaca ccttgatctc agattcctga ctcccagaac 3180

tctgaaaaaa tttttgttgt tttaagccac atttgtgata ttttgttacg gcagccacag 3240

cacggatata gctggtaaca gagttcttta gcctccaaaa tacaagcctg gggtaaataa 3300

ctattgaatt tggccacatg gaaatcagga agaatgtaat aagcatcatc gatggacggg 3360

atgacctggg agatcgtggc aagtttaaag ctaattggaa gttggcaacc aaaaaatcct 3420

gtcttgcaaa ttgccaggaa attgccaaga aaacttttag aaaacatggt taaagatgga 3480

gaaacctgga tgaatccctg gtttctgaag aggtgcttag gcattgaagt gagactgatg 3540

agacttcaca ctgtaccttc agaagggcag acactcacga gatcatcaac cacggctccc 3600

tcgctgggag gcagacacag agctggaggc ggcggccaca aactaggtgg gcagagaggc 3660

gtcagcatcg ggacttgcgg gaggccctgg gaaggaacag gcagcggtgg ggcctggaca 3720

gccctgggga aaggaagtta cttggaggag accaggggct tgcatttggg ccaaaaccaa 3780

ggacaagaga tcagctggtc ccagggaact ctggagccca gcaccccacc acgggtgcag 3840

ggtcctgaca acccgagagg gctgtgggca ccgggcacca gcctgtgggg attctgaagg 3900

ctgctgccca gccacacgtc ccggttctga acaaagggtt caagagacac ctgcaaacct 3960

acctttctga gagcagcctc ttagcctcag ctggtattgg cctttggtca ccccacatgc 4020

cccgacagtg gggcctggcc tcagaaaggg gcccctccat ttgtactttc tatctgatcc 4080

ttgacacagt gctaacacca aagaccaaac ccgaatcttg gttcacatac tctgcaaaga 4140

aatgactaag actagttctt ccggaggtca gttttagaac gtttattctg acttaattct 4200

gccttatctc tgtgcaccaa gaaaactatc tgcacatctg tagtaaaatg tgaaggtacc 4260

ttcattttgt ttgtatacta aacttttaag aaatacctca ctttttaaaa cttggttttg 4320

tgttttgtat gcttataata ttttccataa tagatcattt taatgatgaa ctctgagtct 4380

tctgacagaa tgcagatcaa tggattccta gccatcaggg aggcgttgat tgcaaagggg 4440

cctggtggaa tttttggaga gattctacat cttcttgcaa ctgttcatct ggattacggt 4500

ggtggttgca tgactgtgca tttgttgaaa cttacatgtc acaggatgga ttatactgaa 4560

tgtaaactac acttgagtaa atatgacttt taaaacttta aaaagttggg ggatggttca 4620

ctgccccggc tcgaagcccc ctggccacgc tgcctggcca gcccaccccc atccctgcca 4680

gcgcttgcct ccagcgtccc aacagcctcc tgaccccctg ggctgctcta aaccctcagg 4740

agcgcagcca tccgggatca gctggatgga gatggggagc ccgagactcg tgccacacca 4800

cgtcctcccg cccacaccag ccacacgcag acgtcaaagc agcactgtcc tcgcacgcac 4860

tgctcccacc caccctggca cggccattcc aggcctgggg caggaaggca gatgctcccc 4920

ctgcccccag acacaagcat tcctgcacac acccccagca cacacacgca ctcccatgcg 4980

cacactgaca cacacaggtg tgtgcagctg agacacagcc cgttcccagg aagcccagcc 5040

cccatcactg agggaaaggg gcagcaccgt agggccacag gagtggcagc tggacacaga 5100

gccaggagct ggtgaaggcc caggccactg aggcgggctg ccggcatggc tgggcgtgaa 5160

ggccagaaga gggcaggagg ggctgggggc actatgccta ttgggcctag gtgggcacac 5220

gccgggcagg agaggaacag cccagcccct cagacaggaa ggggtggggg caggggccat 5280

ttgtggaggc cagggcaggg ccagcacccc aaggaaaagc agagcagggt gagaacggac 5340

atggggctca gagctgagca ggcctgctgg gccccaggag ggagacacag atgaccggag 5400

atctcaaggc tggcagaggc cagagatgga gccccagctg ggaagccatc ctccttcctg 5460

ggggcccacg ctgcccggcc cctccagccc agcaagcttg gggcattgga tagaaccggg 5520

agagagccga ccaggcactg aggcccctgc cccaaatgcc cacagcctgg ggaaaatgag 5580

caggtacatg ggaggggcaa gtggagcccc aggcacaccc acacagtgca cacggcctca 5640

cctgggccgg agggggcagg aggctcgcca ccccgctgtg gtttctctcc taatctcacc 5700

ctgggtttct ctcacacttg atgcagatga tgtttctctg acattgtgga ctaagagttg 5760

gtgctggaag gggttagcca tcttggagat gttgctatgg gatgcaggga ttttgcgtgt 5820

gagaaggaca tgattatggg gggaacggag ggcaaactgt catgggttaa aatgtgtccc 5880

ctataaattc atgtgttgaa gtcctaaccc ccaggaccgc agaatgtgac cttgtctgga 5940

aacagtcttt gcagctgcaa tcaagttcag atgaggtcac cctggagtag ggcaagcctc 6000

tgatccaata tgactgctgt cctcatgaaa agggggaatc tgggcacaga cagcacgtgg 6060

ggagaacgcc ctgtgaagat ggtgctgctt ccataagcca agagcaccag agacggccgg 6120

caaagcccag cagcaaggag agagcctggg acagagtctc ccatgacaca gaggagccag 6180

ccccaccgag gcctccatcc cagatgcccg gcctccagaa ccaggacgga ataaacgtct 6240

gttgtttaag ccacgcagtc tggggtgcag tgttgccagg gccacagtta acggatacga 6300

gtgttgtcct gagctgccag ccccacaggc tgcacgaggc ctccctgccc cagcccagtg 6360

cagactcccc agccccctgg gtgtgccatg ggcagtgggg ggcccctcac tccgtcctcc 6420

cccagcctgg gaggttgagc ccattatgag ctccatgggg tgaagctgga acgagaggct 6480

gggagccgac tgggagcctg cggctggagg atggatttcc ccagggaccc acacgtgcac 6540

ctccacctgt ctcctggaca ttctctctga gggcagggct ggtgccagct cagggatcca 6600

gcagggtcac aagggcaggc cgggtccttg tggagagcac atttagtggg agggacatga 6660

tttcccttca aagtgcccat tctggacgct tcccgttcca tgctggacgc ttcctcttcc 6720

acgctggatg cttcctgttc cacgctggat gcttcctgtt ccacgctgga tgtttcctgt 6780

tacactctgg atgcttcctg ttccacactg gatgcttcct gttccatcct ggatgcttcc 6840

tgttccatgc tggacatttc ctgttccact ctggatgctc cctgttccat gctggatgct 6900

tcctgttcca tgctggatgc ttcctgttcc atgctggaca tttcctgttc cactctgcat 6960

gcttcctgtt ccactctgga tgcttcctgt tccacactgg acgcttcctg ctccacgctg 7020

gacgcttcct gttccatgct ggatgcttcc cgttacattc tggatgcttc ccgttccatg 7080

ctggacgctt cctgttccac gctggacgtt tcttgttcca ctctggatgc ttcctgttcc 7140

acgctggatg cttccttttc cacgctggac acttcctgtt ccgcgctgga cacttcctgc 7200

tccacactgg acgcttcctg ctccaggctg gacgcttcct gttccatgct ggatgcttcc 7260

tgttacattc tggatgcttc ccgttccatg ctggacgctt cctgttccac gctggacgtt 7320

tcttgttcca ctctggatgc ttcctgttcc acgctggacg cttcccattc cactctggat 7380

gcttcctgtt ccatgctgga catttcttgt tccactctgg atgcttcctg ttccatgctg 7440

gatgcttcct gttccatgct ggatgcttcc tgttccatgc tggacgtttc ttgttccact 7500

ctggatgctt cctgttacat gctggatgct tcctgttcca tgctggacgt ttcttgttcc 7560

actctggatg cttcctgttc catgctggat gcttcctgtt acattctgga tgcttcctgt 7620

tccatgctgg acatttcctg ttccactctg gatgcttcct gttacattct tgatgcttcc 7680

tgttccatgc tggacatttc ctgttccact ctggatgctt cctgttacat tctggatgct 7740

tcctgttcca tgctggacat ttcctgttcc actctggatg cttcctgtta cattctggat 7800

gcttcctgtt ccatgctgga catttcctgt tccactctgg atgcttcctg ttacattctt 7860

gatgcttcct gttccatgct ggacatttcc tgttccactc tggatgcttc ctgttacatt 7920

ctggatgctt cctgttccac tctggacgct tcccattcca ctctggatgc ttccttttcc 7980

atgctggacc tttcttgttc cactctggat gcttcctgtt ccatgctgga tgcttccttt 8040

tccattccgg acacttccta ttccattctg gacacttcct gtgcgacacc tcctcgggct 8100

tttggtctgc ccagtccctc tggcctcata ccatcccccc ttacctccca cttccacgtt 8160

cgtccttcct cagctcctcc ctctctctag agcttcggcc tggcaaggtc cctcctgatc 8220

tcagtccagg ctcccccagc acaggtagga gacttgcacc tgcccttgga cctccccacc 8280

ctgcatgatg ccagcatccc ccaggcccca gggaggcccc atttctctct ctgcttgtag 8340

tccagtggcc ctggagtgcc actgcaactc gggtgtgccc ctcgcctctg aggaagctaa 8400

gtgccctaag ctaagcagag gccatcccct ctgctcagcc ccagggccct gccccctacc 8460

ccttcccctc acctgcacca caggctctgg ccaactctgc ccaggctctg aatgggcccc 8520

tctggctccc ctctgctgct acactgccct gcaccacctc cactcagctt cagtgtgttc 8580

atccacctgt cccacgtccc ctcggccccc aggagcacag ctggtggccc tggctcctgg 8640

cagcccatct tgttccttct ggagcaccag cctcagaagc cttcctgtgc agggtccact 8700

cggccagccc tgggaccctc ctggtctcaa gcacacacat tctccctgca gccagacctg 8760

cccctgcctg tgagctcaga cctgagcctt ggaacgcctt cccttctcca tcccagctcg 8820

cctttgccag ctgctcagcg ggatgaactc acactcccct ccctgcacca tgagtgagag 8880

ccagctggag agacgcccag gccaaagcag ccaccagggc ccagtggggg tcagaagctt 8940

caggtgagag gcccaggtat tgagaggctg agaccacggg cagaatggtc ataatcactg 9000

ccagtatcag tccagcccca gggactcaga gacagagaaa agagcagtga acaaggtccg 9060

ggctccccac cttctcccac gagtatgggg gcagccacca cccccatccc cacacaccca 9120

tgaggcagcc tcggctgtgt ctggactccc ccttaccctg tgacacagaa accaccagaa 9180

gaaaagggaa cttcaggaag taagcggtgc cgccggtttc aatcctgttc ttagtctttg 9240

cagcgtggag ttcacacccc tggggacctg agggccgagc tgtgatttcc taggaagaca 9300

aatagcagct gacggcgtgg gcaagtctgc ccacatgtac cgcgccaaaa caggaagggc 9360

tgagaccccc acctcggtga gtagggtcag cacagggcaa gggcacaggc tcgggaggag 9420

aaggacagag cctgggtgca gccgtgggcg ctcctggacc tcagctgctg aacaggctac 9480

aagaggctgg ggagacgtgg gggcaaggcc agccccacat ggagacccaa gcggagccag 9540

cacgggggag gtgggcagcc ttcaggcacc aacgcccacc cagtgcaaga tgacggggac 9600

cgtgggcagg ggcttccaag ccaacagggc aggacacacc agaggctgac tgaggcctcc 9660

atgacgacca ggctgggagc acgaggaacc tgacgggatg cggcagagcc ggccgtgggg 9720

tgatgccagc atgggcagga cccacctgag ctgaggaggc agtagaacga gggaggagga 9780

gaggccccag gtgaacggag gggcttgtcc aggccagcag catcactgga gcccagggca 9840

gggtcagcag tgctggccgt ggggccctct ctcagccagg accaaggaca gcaggtgagc 9900

cgggagcaga gcagggaggg tgagtgtggc agcaggacag gagggtggaa gccaaggagc 9960

ccagaggcag aggcagggac aggggaggca caggggctag gctcagagcc acctgatggc 10020

gctggggcac ctgctggcgg ggagcagggc tgtggtcagc agcggagtgg aggggagagc 10080

tgtgctgagt gcacagatgg gaggagggaa gagtccaggg aggcccagaa aggcccagag 10140

tgcagcaggc ctggggcgag gggaggggtg aggctccgtg cgttcaggga gctgacccag 10200

cagagcagag gccactgagg agctgaggtt ctggagaggc ttccagagca ggagcagtgc 10260

agggacggga ggatctggga gctcacccag gaggggcaca tgggcaaggg caaggggctc 10320

tgttggggag acctgactgg acactggggc tgctccacag catagggaac aagccaagtg 10380

ctgcaaaaac aaaaatgagg ccagaaaaac agcccaaacc tggacagagg gtgccaggac 10440

aggcaggggg gcaacagtga cctgagtgac attgctgccc gggttgaggg agggcagagt 10500

gagcaggggg caggcattgg agttcagggt accaggaccg agcagccaca ggtgagcagg 10560

gcaggtgggg gcagaaggag cagggggcac ctcctggagc tcagcagacc agggcagagc 10620

aactgaaggt gaacaagggc aggtgggagg caggatgagc agggggaaga ccctggagct 10680

caggggacca gggcagagca gcctcaggtg cctcaggtga gcaggggctg gtgggtggca 10740

ggacgagtag gggacagctc ctggagctca ggggaccagg acagagcatc aagagctgag 10800

catggctagt gggaggtggg cgaacagggt gcagcccctg gaactcggga ccagggcaga 10860

gcagcggcag gtgagcacgg gctggtggga ggcaggagga acagggggca gctctgggac 10920

ttcaggggac caggggaggg catctgaagg tgaacagggg ctggtggggg caggaagagc 10980

agggggaagc ccctggagct caggggacca gggcagagca gccacaggtg agcaggggct 11040

ggtaggaagc aggaggagca ggggacagcc cctggagctc agagcaccag ggcagagcac 11100

cctcaggtaa gcaggggcag gtaggaggca ggacgagcag gggacagccc ctggagctca 11160

ggggacagag gagagcatca gaaggtgagc aggactgagg cttagcctca gggaatcaga 11220

gcagagcagc cacaggtgag cagggccggt gggaggcagg acgagcaggg gacaggcact 11280

agagctcagg gcaaggcaac cacaggtgag cagggccggt gggaggcatc actcagctcc 11340

tagattttgg caggagctgg gtagttgctg gcagcagaca gctgagggct ggtgaaagtg 11400

cagtgcagcc tcctggtgcc aggaagggag tgtgagccca tcccactgag cagttggcaa 11460

gggtgagctg ggatggagaa gggaaggcat tccagggctc ggggctgagc tctcaggcag 11520

gggcaggtgt ggctgcaggg ggaatgtgtg cttgagacca ggagggtccc agggctggcc 11580

ccagcggacc ctaggcagga aggcctctga ggctggcgcc ccagaaggag caagatgggc 11640

tgccaggagc cagggccacc agcacaatga agctgagtgg aggtggtgca gggcagtgta 11700

gcagcagagg gctgccagag gggcccattc agggcctggg cagagtcagc cagagcctgt 11760

ggtgcaggtg aggggaaggg gtggtgagcg gggccctggg gctgagcaga ggggatggcc 11820

tggctgaggg cagggcactt agcctcctca gaggtcaggg gcacacccca cctgcagtgg 11880

gactccaggg ccactgggcc agcggcagag agaaatgggg cctccctgtg gcctgggggt 11940

cctggcacca cgcagggtgg ggagggccaa gggcaggtgc aaggctccta cctgtgctgg 12000

ggggcctggg ttgagcccag cagggacctt gccgggggaa gctctggaga gagggaggag 12060

gtgggctggt ggccgagaag gccaggccag ggctgggagg gtgaggttgt ggtgactgag 12120

cctccagaag taatgcagga cactgggagg cagggggcat ccaggcactc agggccctga 12180

cctgggctgc tgcacactgg ggctaagggg aaaggagggg agaggctgag gaggaggctc 12240

caggaggcta ttccaaggca gggggttccg gggccctggg gctgaagggc gccgacccta 12300

tgcagtgtct ggcccctctg ctgcacagaa gaaaagggcc ttggagggca gagggcaggc 12360

tatgaccagg gccctgggca agtcaggccc actcactagc ggagggccac gctggggcgg 12420

cagggtcagg agcttcaggg gactcggggg acccacgaga agccatctga gaacagtgtc 12480

cactggtcaa gccaggcacc cataaaaggc tggagtgggg ccaatgggca tgagccgtcc 12540

ctgaggtggc accgatggcc agagctgagg ccaagctaga ggccctggac tgtgctgact 12600

cccggcaggc acagagcgct gacctggctg ccgagccccg cctcctaggc tgcaggggtg 12660

cctgcagaag ggcaccacag ggccaccggt cctgcaagct ttctggggca ggccgggcct 12720

gactttggct ttggggcagg gagggggcta aggtgacgca ggtggcgcca gccaggtgca 12780

cacccaatgc ccgtgagccc agacactgga ccctgcctgg accctcgcag atagacaaga 12840

accgaggggc ctctgcgccc tgggcccagc tctgtcccac accgcggtca catggcacca 12900

cctctcttgc agcctccacc aagggcccat cggtcttccc cctggcgccc tgctccagga 12960

gcacctccga gagcacagcg gccctgggct gcctggtcaa ggactacttc cccgaaccgg 13020

tgacggtgtc gtggaactca ggcgctctga ccagcggcgt gcacaccttc ccggctgtcc 13080

tacagtcctc aggactctac tccctcagca gcgtggtgac cgtgccctcc agcaacttcg 13140

gcacccagac ctacacctgc aacgtagatc acaagcccag caacaccaag gtggacaaga 13200

cagttggtga gaggccagct cagggaggga gggtgtctgc tggaagccag gctcagccct 13260

cctgcctgga cgcaccccgg ctgtgcagcc ccagcccagg gcagcaaggc aggccccatc 13320

tgtctcctca cccggaggcc tctgcccgcc ccactcatgc tcagggagag ggtcttctgg 13380

ctttttccac caggctccag gcaggcacag gctgggtgcc cctaccccag gcccttcaca 13440

cacaggggca ggtgcttggc tcagacctgc caaaagccat atccgggagg accctgcccc 13500

tgacctaagc cgaccccaaa ggccaaactg tccactccct cagctcggac accttctctc 13560

ctcccagatc cgagtaactc ccaatcttct ctctgcagag cgcaaatgtt gtgtcgagtg 13620

cccaccgtgc ccaggtaagc cagcccaggc ctcgccctcc agctcaaggc gggacaggtg 13680

ccctagagta gcctgcatcc agggacagac cccagctggg tgctgacacg tccacctcca 13740

tctcttcctc agcaccacct gtggcaggac cgtcagtctt cctcttcccc ccaaaaccca 13800

aggacaccct catgatctcc cggacccctg aggtcacgtg cgtggtggtg gacgtgagcc 13860

acgaagaccc cgaggtccag ttcaactggt acgtggacgg cgtggaggtg cataatgcca 13920

agacaaagcc acgggaggag cagttcaaca gcacgttccg tgtggtcagc gtcctcaccg 13980

tcgtgcacca ggactggctg aacggcaagg agtacaagtg caaggtctcc aacaaaggcc 14040

tcccagcccc catcgagaaa accatctcca aaaccaaagg tgggacccgc ggggtatgag 14100

ggccacatgg acagaggccg gctcggccca ccctctgccc tgggagtgac cgctgtgcca 14160

acctctgtcc ctacagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 14220

gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctaccccagc 14280

gacatctccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacacct 14340

cccatgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 14400

aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 14460

tacacacaga agagcctctc cctgtctccg ggtaaatgag tgccacggcc ggcaagcccc 14520

cgctccccag gctctcgggg tcgcgcgagg atgcttggca cgtaccccgt ctacatactt 14580

cccgggcacc cagcatggaa ataaagcacc cagcgctgcc ctgggcccct gcgagactgt 14640

gatggttctt tccgtgggtc aggccgagtc tgaggcctga gtggcatgag ggaggcagag 14700

cgggttccac tgtccccaca ctggcccagg ctgtgcaggt gtgcctgggc cgcctagggt 14760

ggggctcagc caggggctgc cctcggcagg gtgggggatt tgccagcgtg gccctccctc 14820

cagcagcagc tgccctgggc tgggccacgg gaagccctag gagcccctgg ggacagacac 14880

acagcccctg cctctgtagg agactgtcct gtcctgtgag cgccctgtcc tccgacctcc 14940

atgcccactc gggggcatgc ctagtccatg tgcgtaggga caggccctcc ctcacccatc 15000

tacccccacg gcactaaccc ctggctgccc tgcccagcct cgcacccgca tggggacaca 15060

accgactccg gggacatgca ctctcgggcc ctgtggaggg actggtccag atgcccacac 15120

acacactcag cccagacccg ttcaacaaac cccgcgctga ggttggccgg ccacacggcc 15180

accacacaca cacgtgcacg cctcacacac ggagcctcac ccgggcgaac cgcacagcac 15240

ccagaccaga gcaaggtcct cgcacacgtg aacactcctc agacacaggc ccccacgagc 15300

cccacgcggc acctcaaggc ccacgagccg ctcggcagct tctccacatg ctgacctgct 15360

cagacaaacc cagccctcct ctcacaaggt gcccctgcag ccgccacaca cacacaggcc 15420

cccacacaca ggggaacaca cgccacgtcg cgtccctggc actggcccac ttcccaatgc 15480

cgcccttccc tgcagctgag gtcacatgag gtgtgggctt caccatcctc ctgccctctg 15540

ggcctcaggg agggacacag gagatgggga gcgggtcctg ctgagggcca ggtcgctatc 15600

tagggctggg tgtctggctg agtcccgggg ccaaagctgg tgcccagggc aggcagctgt 15660

ggggagctga cctcaggaca ctgttggccc atcccggccg ggccctacat cctgggtcct 15720

gccacagagg gaatcacccc cagaggcccg agcccagcag gacacagtat tgaccaccca 15780

cttcctgtcc agagctgcaa ctggaggaga gctgtgcgga ggcgcaggac ggggagctgg 15840

acgggctgtg gaccaccatc accatcttca tcacactctt cctgctaagc gtgtgctaca 15900

gtgccaccat caccttcttc aaggttggcc gcacgttgtc cccagctgtc cttgacattg 15960

tcccccatgc tgtcacacac tgtccccatg ctgtccccac atgtccctga cactgtcccc 16020

catgctgtcc ccacctgtcc cggacactct cctccgcgct gtcttgacct gtgcccaaca 16080

ctgtccccca cgctatcccc ccatccccaa caatgtcccc cacagtttcc tcctgtcccc 16140

tatccccgac actgtcctcc acactgtccc cacctctccc tgtcactgtc gcccatgctg 16200

cccccacctg tcccaacact ttcctccaag ctgtcctcac ctgtccccaa cactctcccc 16260

cacactctct ccacctgtcc ctgacactct cccccatgct gtccccacct gtccctgatg 16320

ctgtcctcca cactgtcccc acctctccct gtcactgtcc ccatgctgtc ccctgtccct 16380

cacactttcc tccatgctgt cctcacctgt ccccaacact ctcccccact gtctccacct 16440

gtccctgaca ctgtccccca cactgtcccc acctgtccct gatgctgtcg tctgtgctgt 16500

ccacatactg ttggtgacct ggctctgttc tccaagttca agcctcagag caggcagtgg 16560

tgaggccgtg gcacctgggt ggcctgaggg gtgggcgggc cttgggggca gggctgtggc 16620

ctcgctcacc cctgtgctgt gccttgccta caggtgaagt ggatcttctc ctcagtggtg 16680

gacctgaagc agaccatcgt ccccgactac aggaacatga tcaggcaggg ggcctagggc 16740

caccctctgt ggggtgtcca gggccgccca gaccccacac aggagccgtg ggccatgctc 16800

agccatcacc caggccacac ctgcccccga cctcaccgcc ctcaacccca tggctctctg 16860

gcctcgcagt cgccctctga ccctgacacg ccccccttcc agaccctgtg catagcaggt 16920

ctaccccaga cctccgctgc ttggtgcatg cagggcgctg ggggccaagt gtcccctcag 16980

caggacgtcc ctgccctccg gcccgccagg tgctcacaca aaaggaggta gtgaccagca 17040

tcccaggccc ccactcaggc aggacctcgc cctggagcca accctgtcca cgccagcctc 17100

ctgaacacag gcgtggtttc cagatggtga gtgggagcat cagtcgccaa ggtagggaag 17160

tcacagcacc atcaggccct gttggggagg cttccgagag ctgcgaaggc tcactcagac 17220

ggccttcctc ccagcccgca gccagccagc ctccattcca ggcactcccg tgaactcctg 17280

acatgaggaa tgaggttgtt ctgatttcaa gcaaagaacg ctgctctctg gctcctggga 17340

acagtctcag tgccagcacc accccttggc tgcctgccca cactgctgga ttctcgggtg 17400

gaactcgacc cgcagggaca gccagcccca gagtccgcac tggggagaga aggggccagg 17460

cccaggacac tgccacctac cacccactcc agtccaccga gatcactcgg agaagagcct 17520

gggccatgtg gccgctgcag gagccccacg gtgcaagggt gaggatagcc caaggaaggg 17580

ctgggcatct gcccagacag gcctcccaga gaaggctggt gaccaggtcc caggcgggca 17640

agactcagcc ttggtggggc ctgaggacag aggaggccca ggagcatcgg ggagagaggt 17700

ggagggacac cgggagagcc aggagcgtgg acacagccag aactcatcac agaggctggc 17760

gtccagcccc gggtcacgtg cagcaggaac aagcagccac tctgggggca ccaggtggag 17820

aggcaagacg acaaagaggg tgcccgtgtt cttgtgaaag cggggctgct ggccacgagt 17880

gctggacaga ggcccccacg ctctgctgcc cccatcacgc cgttccgtga ctgtcacgca 17940

gaatccgcag acagggagac tcgagcggga gtgcggccag cgcctgcctc agctgtcagg 18000

gaggactccc gggctcactc gaaggaggtg ccaccatttc agctttggta gcttttcttc 18060

ttcttttaaa ttttctaaag ctcattaatt gtctttgatg tttcttttgt gatgacaata 18120

aaatatcctt tttaagtctt gtacttcgtg atgggagccg ccttcctgtg tccacgcgcc 18180

tcctgccccc ggtgggaagc acggtcagga ggaggctggt ccagctgcac ctcgggggct 18240

ccctgcatac gccccccgcc tcctgcagcc acacgcattg cccgagcgac cctccctggc 18300

ccctgtcgct acatggaccc ccggggtttc tcctcttttc tacatggatg cagtttctcc 18360

tcctgctggg cacggtgctg cctgccctgg tcactctgcg ggggacaggg cctccaggga 18420

aagctgggtc gaggctggga gctggctcag gctgcccagg cagagccaca gggagggcct 18480

tccagaacca accatggtcc gaagcgagag gtgggtgtca gatctgtgtg agtcagctca 18540

ggaccacagc ggggcggctc ccacagcaga catggatcct cccaggccta gagaccagga 18600

atctgagatc aggatgcagg cagggctggt ttctctcaag ccctctctcc ttggcttgta 18660

gacaccgtct cctccctggt cctcacatgg ccatccctct gtgtgcccgt gtcctaagct 18720

tctcttctta taagaacaca catcggatta gattagtgac cccctatgaa cttaatgacc 18780

tctgtaaaga ccccatctcc aaatagtcac attgtgaggc cagggattaa gacttgaata 18840

tatgaatttg taggggccac gatttaaccc atcacagtcc agactctggc ccccaaaatt 18900

catgttcttc tcacatgcaa aacacattca tcctgtctca gcatccccct gggcactagg 18960

tcatgtagca aggacggatt ttcaacagaa ataactattg caacagaaga aagagtccgg 19020

catgacctgg actcaccttc atctgtgcag aggccacagc cttgtaaagg gaggtggtag 19080

ggggagcagg gagggtgctc ggggctcagt cgtcggggaa gggaaaagtt gcccagcgct 19140

ggtcagcgtc cccgggatgg gacccgctgt gtctgtgccg gccactgttg aggtcaggat 19200

tctgtcctcc cagagcctgg agacacaggc cccatccttc ccaatgggga cacttcaggg 19260

agtggctctc aggtcccgag aaagaccctc ctgggtcaca ggaaatgcac agacatcggg 19320

aacggataga aggtcgtgtg gttgcggccc tctcagcaga taccctgaga aagggaggtc 19380

ggggttggtc caaacggtga gttctggtgc acggagcttt ctcaggcagg tgttgacggg 19440

gcaggggtcg gcctaggggt acggccagaa gctgttagaa actgttagtg tctgctcaag 19500

tctttacaag ccaaggttga ggccgagtgg agaggctccg aggagcctgg ctggaactca 19560

gtcaaggaca gggtcttgtt actgcagtgg ctgcggtggc tgcggtggct gcgaaatgcc 19620

gtcggagttg cctgtggcag gagagagacc atctcaccca ggaaggagga gtggttggat 19680

tcgtttgtgt ggcatcgagc agctggagct tcaccaaaca cagagttggg gactaaatcc 19740

ccagactcca ggccctgcca tgccgtggga aggctcgcca ctggagggtg cgctccaggg 19800

ggcctggcct gaactgggtg ctgaagccca gccctttaac tctcaggaca cgctgctgca 19860

gccccgcggg gggtgaggga gagagcacct ggggtgcagg gcgggcagct gctgcatcac 19920

cggctctatc ccaagcccaa ggatggcgtc ccagagatgc aggagagctt tgtccagaga 19980

aggtgccagc cctcagggac cctgctggag agatctccac cctctgccct tcaaggggcc 20040

ctacgggcct ccaggtgctc tggtggggtg ggctccagtc cactgtctga ggatggacgg 20100

cctggccagg ataaggaaag gaaacccagg acggtgccgg gctccgggtc attccgtgca 20160

ctgagcaggc tgagttggga agaagcagat gcttcctgca gctgctgccc cctgcagggc 20220

ctggcgcctg gaccaggttc ccctggggaa attgggcccc tccctgagcc acccggggcc 20280

cacctcccac tttctacctg ggaccgagca tcctccagag ggtcagccct cctgcgggaa 20340

caccatgccc agccccagga ccctccctca actctccagc aaggctgccc ctgcacaccc 20400

cccagcagcc catgctgtga tgtaacatga catcgtgtga catggtgtga tgtcctatca 20460

cagtgtgaca tccctggtgt gatgtggtgt gacatggtgg gatgtggtgt gacatggtgt 20520

gatgtccctg gtgtgaggtg gtgttggaca tagtgtgatg tgatgcgaca gatgtaacat 20580

ccctggtgtg atttggtgtg acgtagtgtg atgggtatga catccctggc ttgatgaggt 20640

ataatatgaa atgatgtgga atggtgtgac atgatgtgat gtgatgggac agggtgtgac 20700

gtccctggtg tgatgtgatg taacaggctg tgacatagtg tgatgtgggt ggtgtgacat 20760

gatgtgatgt ggtgtgatgg tgtgggacat ccctggtgtg atggggtgtg attaaacctt 20820

gattccatgc agcacatgtt tctgtgagca cagggttggg gctaaagtta caggttaaca 20880

gcatctcaaa gcagaacaat ttttctttgt acagatcaaa atggagtttc ttatgtgttc 20940

cttttctaca taggcacagt aacagtctga tctttctttt ccccacagtg tgacatggtg 21000

tgacatttct ggtgtgactc ttgtgttgtg acatttgtgg tcaccccagg atacagaggt 21060

ctctgtggcc aagggaaggg ggagaatgga accatctgag catgttgacc tggaggaatt 21120

ggtggccctt gagtccacga agcccaccct tccaggtgcc cctgccccac gtgacccaag 21180

tgggcttgca gagcagcaag caggactctg gttagacagg aggaaggacc tgccaccacg 21240

tggccttgtg aggagacaga gcgaggctgt gacctcggcg tccgcccagc acagggtgct 21300

gctgaagctc ctccggtcct ctcagcagcg gtctcagagc aaggcccaag gcaggctgaa 21360

gagaggggca gagggaggat gctggggagg caggtgtgag gggactgaga gcccaggttt 21420

cagctgagcc cctccacagg gaaggagcct agctgaacac ccatctcccc acacgctccc 21480

aaccctgcct ctgcccgacc acctcccaga gggcacctcg aaccctctgc tacccacact 21540

cagcaagggg tatggtgccc ccaccaagcc cagccagcaa agcctggcac agccacgcct 21600

gtgcccacca ctcccatggc caaggtcact gctaacatgg caggacagag ccaggcctgg 21660

aggagacaga acatcagtcc catggggaag ctccctgctc acacggcagg gccaggcctg 21720

gagaagacag aacaccccat ctggcatggt actcaggctg cacatgcctg ccacgaacgg 21780

gggccacgcg acaatgcctg ccacacatgg gggccacgca acgatgcctg ccacacatgg 21840

gggccacgcg acgatgcctg ccacacatgg gggccacgcg acgatgcctg ccacgcatgg 21900

gggacacgtg acacacacat acacacacgg gcctcacagg cacacaaatg cttgcgaacc 21960

cagcacccac ccagcacact caggcatagg ctccctgggc aggtcacaac ctcacgcctt 22020

gagctagtcc atgtgccagg cccgtcaccc acgtactcgt ccccggtgct agctattgag 22080

ccacaccgtc ttctctgtgg atccctccca gcccactcag cacaatggac atgctctctc 22140

ccgtccagtg actgcaccgg cctcttcccc tcagcaccca agtctggcca tctcccagga 22200

aacccggacc accacgggca gggaccacgt tcctcactgt ccacgtggac caccccacac 22260

ctgaccccag atgcagtcac actacgtcct gcttcaatat tgaaaagggg aaaagctgga 22320

ggagggtaaa gatgaaagag aaaaaagcaa gaggggaggg tcacattctt ctgaggcttt 22380

gattacatct cactgagccc ccacgttgca tgaaaaggag gggtggaggg agcaattaca 22440

catttgcctt gtgctcagta aatctgcact ttataagcaa ataaacagag tagaggaaga 22500

agtcaaatat gcattcgtct caggggcagg agggatgatt tcttgtctca ttttgtccca 22560

tgtcatgaag accgggctgt taatttatat tgtcagggtg agggaggcca cctgggtaga 22620

cctggcctat ctgctgctgc tctcagtttg gaaacaaaag gaaacgcatg actttttttt 22680

tttttttcat gactcagctt cccacctcaa ctgttctttt tggcatagtg actttagggt 22740

cctgagattt tattttcctt tcacaatggt caagatcaca ccctcagtat tcaggagagc 22800

tggtccaacc cagacccctg ccgtccctgg acgcttttca aatacgtgct gcagtctctg 22860

acactctctc accatggtag aaactgaggt cttggtgagt ctgcagcccc tgctcatggg 22920

gacaatgcag gctgaaaccc gcatttctga ccccaaagct cctgttcctt tcactcaccc 22980

ccacaccagc tctttgagtc cagagctttg tccttgcctg agtcctaccc tcagggacag 23040

gggcccaacc cagccaccaa cacatcatac cctgagaggg tgccaggagc ccagagatgt 23100

ttggagagca cagtagccct ggaagctctg tggagatgct gcacatttct ctattcaaca 23160

gatactcgcc aggtggccag cagcagagat gccacatggc atggagctcg ccttggccaa 23220

caggacaggt gtggggttgg gaggccttcc cagggcactc cttgaagcag agctgtgggg 23280

aaaagttggg gccaatctca gatctccccg tcaacaccgg gtctcctgcc ctcctgggcc 23340

acagaaaact aagctccgtg gatactgcgg ctgggtgggg ccgtggggga gaagaaatca 23400

caacgagtta aaagatcatt ttctaaaact gttatgatca ggactcacat aaccatatga 23460

cgacacattt cagagatgct ctttatctca ttaattaagg tgtcgtaacc agttcaaagt 23520

ggaattctaa gtactacact tacataattg attcaggaat gctaaaagga gttcatagat 23580

agatgcaaaa ctggcctttt ccctggaaga tgaggagcaa ttcattgtcc ttccaaagat 23640

gagaacttga atttctacca actcaaagag cttttgcatt gctatcaatt atgtacaact 23700

tagagcagtg gtccccaaca tttttggcac caggaaccag ttccatggaa gacaattttt 23760

ccacagacca ggatcggggg atggcttggg gacaaagctg ttccacctca gatcattagg 23820

cattagggtg tcataaggag tgtgcaactt agatcccggg aatgtgcggc tcgcaatagg 23880

gttcgctcct gtgagaatct aatgctgcca ctgatctgac aggaggtgga gctcgggcag 23940

gaatgctcac acacccctca cctcctgctc tgtggcccag ttcctaacag gccatgaacc 24000

ggttccagtg catgacccag ggattgggga cccctggctt atagaggtgt aaaatagttc 24060

aaaggaaata aaagatgcag agctccacag aatgaaataa cctggaagag tgtacaagac 24120

gatgccttgc tttccatgga aggcacctac taatttttct caatgtttcc tataaacata 24180

tataactgac tgacagaaac agatccataa tataaagaag acccctgtaa accaatgaga 24240

aaaaaaatca aataatccaa cgaggaatag gcaagagaat tgaacagatg ttttacagaa 24300

gatatccaaa tagccactaa acatatgaaa aggtgttgaa ccacactagt caacagggaa 24360

atgaaaatga aaaaccacat gagagaaagt agttctgatt ccagtaatgc tagagcagct 24420

aatatcagac tagccctttg gcagatggca attataaaca ctggaaacac tgtaagcaca 24480

cacaacaccc acacacacca attgcaggca ctggaacatg accagaagta ggcaaacact 24540

agtaaggatt attccgtgaa atattcgtct gaagtcacac cccagtgcat gtaatgggtg 24600

cagctagagt ttaagcagga aactgcagcc ctcctggtga ggagtgggat gcagggctgc 24660

attttcagag cagctggaaa tgaagagaag atgtccataa aggagaagct caccgaaggg 24720

aaaccacaca atctgcaagt aaactccact gaaacctctg gccgatccct taggtgtgca 24780

tgggtaggga aaactccaaa gggcccagca gaaagcaaca cctgtaagtc gagagaactg 24840

agattccagc tactgccaac tgccaggcag acagacttgg gagtttgagt caactcaagc 24900

taactgctaa cattgaaaaa aacaattaat gctctgctaa gaaagaatgc aaaccccata 24960

gcctgtacgc atgttatcaa catcaggtgc acatccaaaa ttaccatgga tgcaaagaaa 25020

catgaaaatg tgatccatag tcagaagaaa aagggatcaa tggagatcaa ctccaagatg 25080

acctagatgc agatacagtt atcagacaag gactttaaag aagttatgtt aaacatgttc 25140

aaagacttaa aggaaaatac tattataatg agtgactaga tggggaatct ctataagaga 25200

cggaaaatat ttacaagaac caaatgaaaa ttctagaact gaaagtatga ttctgaaatg 25260

aaaaacatca tttttcagag cagggtgaga atggacatgg gactcagagc tgagcaggcc 25320

tggtgggccc caggagggag acacagagga ctgggggatt tcaaggctgg cagaggccag 25380

agatggatcc ccagctggga ctggacctgg gcttatggga gcaacaggtg acccatcctc 25440

cttcctgggg gcccaccctg cccggcccct ccagcccagc acaggcattg gatagaaccg 25500

ggagagagca ggccaggcac tgaggcctct gccccaaatg cccacagcct ggggaaaatg 25560

agcagataga tgggggggca agtggatccc caggcacacc cacacagtgc acacagcccc 25620

acctgggcca gagggggcag gaggctcgcc acccctgctg tggtttctcc cacacttgat 25680

gcaggtgata ttcctctgag attgtggact aagagttggt gctggaaggg gttagccatc 25740

ttggagatgt tgctatgggg tgcagggatt ttgcatgtga gaaggacatg attatggggg 25800

gagcggaggg caaactgttg tgggttaaaa tgtgtcccct ataaattcat gtgttgaagt 25860

cctaaccccc aggaccacag aatgtgacct tgtttggaaa cagtctttgc aactgcaatc 25920

aagttcagat gaggtcaccc tggagtaggg caagcctctg atccaatatg actgctgtcc 25980

tcatgaaaag ggggaatctg ggtacagaca gcacgtgggg agaacaccct gtgaagatgg 26040

tgctgcttcc ataagccaag agcagcagag acggccggca aagcccagca gcaaggagag 26100

agcctgggac agagtctccc atgacacaga ggtgccagcc ccgccgaggc ctccatccca 26160

gatgcccggc ctccagaacc aggacggaat aaacgtctgt tgtttaagcc acgcagtctg 26220

gggtgctgtg ttgccagggc cacagttaac ggatacgagt gttgtcctga gctgccagcc 26280

ccacaggctg cacgaggcct ccctgcccca gcccagtgca gactccccag ccccctgggt 26340

gtgccatggg cagtgcgggg cccctcactg catcctcccc cagcctggga ggttgagccc 26400

attatgagct ccatggggtg aagccggagc cagaagctgg gagccgactg ggagcctgcg 26460

gctggaggat ggatttcccc agggacccac acgtgcacct ccacctgtct cctggacatt 26520

ctctctgagg gcagggctgg tgtcagctca gggatccagc agggacacaa gggtgggccg 26580

ggtccttgtg gagagcacat ttagtgggag ggacatgatt tcccttcaaa gtgcccattc 26640

tggatgcttc ctggtccacg ctggacactt cctgttccac gctggacgct tcctgttcca 26700

cgctggacgc ttcctgttcc acgcttgatg tttcctgttc catgctggat gcttcctgtt 26760

ccatgctgga catttcctgt tccactctgg atgctccctg ttccattctg gatgcttcct 26820

gttccatgct ggacatttcc tgttccactc tgcatgcttc ctgttccact ctggatgctt 26880

cctgtgcgaa acctcctcgg gcttttggtc tgcccagtcc ctctggctgc atctcgtccc 26940

ccgctacctc ccacctccac atccgtcctt gcccagctcc tctctctctc cagagtttcc 27000

acctggcaag gtccctgatg agctcagtcc aggctccccc agcacaggta ggagcctagc 27060

acctgccctt ggacctcccc accctgcatg atgccagcat ccccaggccc cagggaggcc 27120

ccatttctct ctctactgct ggcccagtgg ccctggagtc ccactgcaac tcgggtgtgc 27180

ccctgacctc tgaggaagtt aagtgtcctg tccctagcca ggctatcccc tctgctcagc 27240

cccagggccc tgccccttac cccttcccct cacctgcacg ataggctctg gccaactctg 27300

cccaggccct gaatgggccc ctctggctcc cctctgctgc tacactgccc tgcaccacct 27360

ccactcagct tcagtgtgtt catccacctg tcccaagtcc cctcggcccc caggagcaca 27420

gctggtggcc ctggttcctg gcagcccatc ttgttccttc tggagcacca gcctcagagg 27480

ccttcctgtg cagggtccac tcggccagcc ctgggaccct cctggtctca agcacacgtt 27540

ctccctgcag ccagacctgc ccctgcctgt gagctcagac ctgagccttg gaacgtcttc 27600

ccttctccat cccagctcgc ctttgccagc tgctcagtgg gatgaactca cactcccctc 27660

cctccaccat gagtgagagt cagctggaga gatgcccagg ccaaagcagc caccagggcc 27720

cagtgggggg ccagaagctt caggtgagag gcccaggtat tgagaggctg agaccatggg 27780

cagaatggtc ataatcgctg ccagtctcag tccagcccca gggactcaga gacagagaaa 27840

agagcagcac acaaggtccg ggctccccac cttctcccgt gagtatgggg gagtatgggg 27900

gcagccacca cccccatccc cacacaccca tgaggcagcc tcggctctgt gtggactccc 27960

cctcgccctc tgacacagaa accaccagaa gaaaagggaa cttcaggaag taaggggtgc 28020

cgctggtttc aatcctgttc ttagtctttg cagcgtggag ttcacacccc tggggacctg 28080

ggacctgagc tgtgatttcc taggaagaca aatagcggct gacggcgggg gcggggccgc 28140

ccacatgtac ctcgccagaa caggaagggt tgagaccccc acctcggtga gtggggtcag 28200

cacagggcag gggcacaggc tcgggaggag gacagcctgg gcgcagccgt cggcgctcct 28260

agacctgagc tgctgaacag gctgcaagag gctggggaga cgcgggcgcg aggccagccc 28320

cacatggaag cccaagcgga gccagcacgg gggaggtggg cagccttcag gcactgatgc 28380

ccacccagtg cgagacgacg gggaccgtgg gcaggggctt ccaagccaac agggcaggac 28440

acaccagagg ctgactgagg cctccaggac gaccgggctg ggagcacgag gaacatgacg 28500

ggatgcggca gaaccggctg tggggtgatg ccaggatggg cacgaccgac ctgagctcag 28560

gaggcagcag agcgagggag gaggagaggc cccaggtgaa cggaggggct tgtccaggcc 28620

ggcagcatca ccagagccca gggcagggtc agcagagctg gccgtagggc cctcctctca 28680

gccaggacca aggacagcag gtgagccggg agcagagcag cgagggtgag tgtggcagca 28740

ggacagaagg gtggaagcca aggagcccag aggcagaggc agggacaggg gagggacagg 28800

ggctgggctc agagccagct gatggggctg gggcacctgc tggcggggag cagggctgtg 28860

gtcagcagcg gagaggaggg gagagctgtg ctgagtgcac gggcgggagg agggaagagt 28920

ccagggaggc ccagaaaggc ccagagtgca gcaggcctgg ggcgagggga ggggctgagg 28980

cccagcagag cagaggccac tgaggagctg aggttccgga gaggcttcca gagcaggagc 29040

agtgcaggga cgggaggatc cgggagctca tccaggaggg gcacataggc aaggggctct 29100

gttggggaga cctgactgga cactggggct gctccacagc atagggaaca agccaagtgc 29160

tgcaaaaaca aaaatgaggc cagaaaaaca gcccaaacct ggacagaggg tgccaggaca 29220

ggcagggggg caacagtggc ctgagtgaca ttgctgcccc gggttgaggg aggacagagt 29280

gagcaggggg caggcattgg agttcagggt accaggaccg agcagccaca ggtgagcagg 29340

gcaggtgggg gtagaaggag cagggggcag ctcctggaac tcaggggacc agggcagagc 29400

agccacaggc aaacaggaga ggggaggggg ggcaggagga gcagggggca gctcttggag 29460

ctcaggggac cagggcagag acgccgcagg tgagcagggg caggtggggg ggcaggagga 29520

gcagggggca gctcttggag ctcaggggac cagggcagag cagccacagg tgagcagggg 29580

caggtggggg gcagaaggag tagggggcag ctcttggagc tcagaggacc agggcagagc 29640

agccacaggg gaggaggggc aggtgggagg caggatgaac agggggcggc tcctggaact 29700

caggaaacag gggagagcat cagaaggtga gcagggccag tgggaggttg cagagcaggg 29760

gacagctcct ggagctcagg ggaccagggc agagccgccg caggtgagca ggggcaggtg 29820

gggggcagga ggagcagggg gcacctcctg gagctcaggc gaccggggca gagcagcctc 29880

aggtgaaaag ggccggtggg gggcaggagg agcaaggggc agctcctgga ggtcagggga 29940

ccagggcaga gccgccgcag gtcagcaggg ccggtgggag gcaggacgag caggggacag 30000

gcactagagc tcagggcaag gcagccacag gtgagcaggg ctggtgggag gcatcactca 30060

gctcctagac tttggcagga gctgggtagt tgccggcagc agacagctga gagctggtga 30120

aagtgcagtg cagcctcctg gtgccgggaa gggagtgtga gtccatccca ctgagcagtt 30180

ggcaagggcg agctgggatg gagaagggaa ggcattccag ggctcagggc tgggctctca 30240

ggcaggggca ggtgtggctg cagggggaac gtgtgcttga gaccaggagg gtcccagggc 30300

tggccccagc ggaccctggg caggaaggcc tctgaggctg gcgccccaga aggagcaaga 30360

tgggctgcca ggagccagga ccatcagcac aatgaagctg agtggaggtg gtgcagggca 30420

gtgtagcagc agagggctgc cagaggggcc cattcagggc ctgggcagag tcagccagag 30480

cctgtggtgc aggtgagggg aaggggtggt gagcggggcc ctggggccga gcagagggga 30540

tggcctggct gagggcaggg cgcttagcct cctcagaggt caggggcaca ccccacctgc 30600

agtgggactc cagggccact gggccagcgg cagagagaaa tggggcctcc ctgtggcctg 30660

ggggtcctgg caccatgcag ggtggggagg gccaagggca ggtgcaaggc tcctacctgt 30720

gctggggggc ctgggttgag cccagcaggg accttgccgg gggaagctct ggagagaggg 30780

aggaggtggg ctggtggccg agaaggccag gccagggctg ggagggtgag gttgtggtga 30840

ctgagcctcc agaagtaatg caggacactg ggaggcaggg ggcatccagg cactcagggc 30900

cctgacctgg gctgctgcac actggggcta aggggaaagg aggggagagg ctgaggagga 30960

ggctccagga ggctattcca aggcaggggg ttccggggcc ctggggctga agggcgccga 31020

ccctatgcag tgtctggccc ctctgctgca cagaagaaaa gggccttgga gggcagaggg 31080

caggctatga ccagggccct gggcaagtca ggcccactca ctagcggagg gccacgctgg 31140

ggcggcaggg tcaggagctt caggggactc gggggaccca cgagaagcca tctgagaaca 31200

gtgtccactg gtcaagccag gcacccataa aaggctggag tggggccaat gggcatgagc 31260

cgtccctgag gtggcaccga tggccagagc tgaggccaag ctagagacac tggactgtgc 31320

tgactcccgg caggcacaga gcgctgacct ggctgccgag ccccgccccc taggctgcag 31380

gggtgcctgc agaagggcac cacagggcca ccggtcctgc aagctttctg gggcaggccg 31440

ggcctgactt tggctggggg cagggagggg gctaaggtga cgcaggtggc gccagccagg 31500

cgcacaccca atgcccgtga gcccagacac tggaccctgc atggaccatc gcagatagac 31560

aagaaccgag gggcctctgc gccctgggcc cagctctgtc ccacaccgcg gtcacatggc 31620

accacctctc ttgcagcttc caccaagggc ccatcggtct tccccctggc gccctgctcc 31680

aggagcacct ccgagagcac agccgccctg ggctgcctgg tcaaggacta cttccccgaa 31740

ccggtgacgg tgtcgtggaa ctcaggcgcc ctgaccagcg gcgtgcacac cttcccggct 31800

gtcctacagt cctcaggact ctactccctc agcagcgtgg tgaccgtgcc ctccagcagc 31860

ttgggcacga agacctacac ctgcaacgta gatcacaagc ccagcaacac caaggtggac 31920

aagagagttg gtgagaggcc agcacaggga gggagggtgt ctgctggaag ccaggctcag 31980

ccctcctgcc tggacgcacc ccggctgtgc agccccagcc cagggcagca aggcaggccc 32040

catctgtctc ctcacccgga ggcctctgac caccccactc atgctcaggg agagggtctt 32100

ctggattttt ccaccaggct ccgggcagcc acaggctgga tgcccctacc ccaggccctg 32160

cgcatacagg ggcaggtgct gcgctcagac ctgccaagag ccatatccgg gaggaccctg 32220

cccctgacct 32230

<210> 70

<211> 326

<212> PRT

<213> Intelligent people

<400> 70

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg

1 5 10 15

Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

20 25 30

Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

35 40 45

Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

50 55 60

Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr

65 70 75 80

Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys

85 90 95

Thr Val Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro

100 105 110

Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp

115 120 125

Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp

130 135 140

Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly

145 150 155 160

Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn

165 170 175

Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Val His Gln Asp Trp

180 185 190

Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro

195 200 205

Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu

210 215 220

Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn

225 230 235 240

Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile

245 250 255

Ser Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr

260 265 270

Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys

275 280 285

Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys

290 295 300

Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu

305 310 315 320

Ser Leu Ser Pro Gly Lys

325

<210> 71

<211> 1901

<212> DNA

<213> Intelligent

<400> 71

gcctgacttt ggctttgggg cagggagggg gctaaggtga ggcaggtggc accagccagg 60

tgcacactca atgcccgtga gcccagacac tggaccctgc ctggaccctc gcggatagac 120

aagaaccgag gggcctctgc accctgggcc cagctctgtc ccacaccgcg gtcacatggc 180

accacctctc ttgcagcctc caccaagggc ccatcggtct tccccctggt gccctcctcc 240

aggagcgtct ctgagggcac agcggccctg ggctgcctgg tcaaggacta cttccccgaa 300

ccggtgacgg tgtcgtggaa ctcaggggcc ctgaccagaa gcgtgcacac cttcccggct 360

gtcctacagt cctcaggact ctactccctc agcagcgtgg tgaccgtgcc ctccagcagc 420

ttgggcaccc agacctacac ctgcaacgta gatcacaagc ccagcaacac caaggtggac 480

aagacagttg gtgagaggcc agcacaggga gggagggtgt ctgctggaag ccaggctcag 540

ccctcttgcc tggacgtacc ccggctgtgc agccccagtc cagggcagca aggcaggccc 600

catctgtctc ctcacccgga ggcctctgcc cgccccactc atgctcaggg agagggtctt 660

ctggcttttt ccaccaggct ccaggcagcc acaggctgga agcccctacc ccaggccctg 720

cgcacaaagg ggcaggtgct gcacttagac tggccaagag ccatatccgg gaagaccctg 780

cccctgacct aagcccaccc caaaggccaa gatctccact ccctcagctc agacacctct 840

cctcccagat ctgagtaact cccaatcttc tctctgcaga gcccaaaacc ccatgttgtg 900

acacaactca cacatgccca ccatgtgcaa gtaagccagc ccaggcctcg ccctccagct 960

caaggcggga caggtgccct agagtagcct gcgtccaggg acaggcccca accgggtgct 1020

gacacgtccg cctccatctc ttcctcagca actgaacccc tggggggacc gtcagtcttc 1080

ctcttccccc caaaacccaa ggataccctc atgatctccc ggacccctga ggtcacgtgc 1140

gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta cgtggacggc 1200

gtggaggtgc ataatgccaa gacaaagccg tgggaggagc agtacaacag cacgtaccat 1260

gtggtcagcg tcctcaccgt cgtgcaccag aactggctga acggcaagga gtacaagtgc 1320

aaggtctcca acaaaggcct cccagccccc atcgagaaaa ccatctccaa aaccaaaggt 1380

gggacccacg gagcgcgaag gccacgtgga cagaggccgg cttggcccac cctctgccct 1440

gggagtgacc gctgtaccaa cctctgtccc tacagggcag ccccgagaac cacaggtgta 1500

caccctgccc ccatcccaga agatgaccaa gaaccaggtc accctgacct gcctggtcaa 1560

aggcttctac cccagcgaca tcgccgtgga gtgggagagc aatgggcagc cggagaacaa 1620

ctacaagacc acgcctccca tgctggactc caacggctcc ttcttcctct atagcaagct 1680

caccgtggac aagagcaggt ggcagcaggg gaacgtcttc tcatgctccg tgatgcatga 1740

gggtctgcag aaccactaca cgcagaagag cctctccctg tccccggggt aaatgagtgc 1800

gacggccggc aagcccccgc tccccgggct ctcgcggtcg cacgaggatg cttggcacgt 1860

accccgtcta catacttccc aggcacccag catggaaata a 1901

<210> 72

<211> 32230

<212> DNA

<213> Intelligent

<400> 72

gaattcacca ttttaaagtg tacaattgag tggcatttag tacattcacg atgttgtgca 60

atcatcactc ctattattac tctgtaaact ttacaaattc atcgtattta tttatttaat 120

ggttgttaca gtttgcaact aaaatagaag gaaatacatc aaaatattag cagtggctct 180

tctgggtgaa gtacaagggg caaattgttt tctcctgttt gtttttctgt acacgttgtt 240

tactgtgtca cattgtttac tgtgaatagt tactatttga aactaaagat caagggcgat 300

aaactgtatt ttaagaggaa gtcagaaaac aaacaaggtg gccctctcgt atgctgtgct 360

gacacctgtg tccctgtagc ctcaactacg cctgttgccc ccacctgcgc ctgaagtttg 420

aggtcgctgg tggcacctct gtgccagggg gccctggaac ctctcctagg atggtcccag 480

agaaaggaga gagatctgct tcctggaatc tcagctctag atgctttggt caactccatg 540

tcatctgtgt ttgataagct tgatttccat ttgttttggc ccctctttta gtgtgctttt 600

tgttagcaga aactcacgaa ctagagaact gcaggtcccc caggggagtg ggttgagatg 660

tttttgacat cagccacctc tttctcccac agttttgtta ttaaaacccc attttaacaa 720

ccttgaaaga caggttttaa aagaaagttg caattttctt acaattctac tgcttaatta 780

aaagagtagg ccctgtggtt ccctttcccc atcctggtgg tgttctccct gcccctgtgg 840

tggtgttctc cctgcccctg cctctcacca gtgtatgctg ttcccaggtg gcagtaatcc 900

tatctcaggt ctggcattcc aaggcaggct cctgctcctg ggtggggagg ggcctccatg 960

ctgggttgaa tgtctgtgct ttgctgtccc ctgacccaca gagtgccctt ggacaggaca 1020

taggtctcag tttcctaagt gagagtcccg gattagatgg actgtggtta accctttatg 1080

tgtcatggcc tctttctgta taaagccaca gatctttccc tgcaatgagc atttatacag 1140

acacttttca gggttttgtg aacccgtgag ctggacatcc tagagctctc cttcaatccc 1200

atggcctcta gggtaccctg ctctggaaag gcaagactgg gcctcttttt tgggaccccg 1260

gagggcagac cttgggtttc gtcgtggcac ccctgctggg tgatgtgccc agggctttcc 1320

acgatgatcc cactgaatcc tcacatagcc ctgaggggaa gctgccacca tcatcacctc 1380

cacatcacag aggagtacac aggcaggaca gtcacactcg gacaagtggt ggagccagga 1440

cacaaaccca ggtctgtctg gactctaagc ttaggctctt accatgaact gtgatccaac 1500

catcgggaga gtcagcccaa tgaccttcaa gttccctcca atgctaaggt cacacctgag 1560

aagcttgcta gggtttctaa agcatcttcg tgcacatatg tccttccagg gttcttccca 1620

gcagtttgtg agatgggcag ggcagatgaa gggaaaaggc ttcaagacat taaggaaagg 1680

gcccaagatc acacccgagt caggatcaga gcaggaattg gaatctaatc tctacacaga 1740

gttgccagat accaatctgt gcttctcatg gttagaattt acctacttat ccaactattc 1800

acccatccaa ctacccaccc acacatgtac ccaccagtcc acgcattcat ctatccagcc 1860

atccactcac ccacccaact atccactcat ccattcacct accaatccac ccactcatcc 1920

acccatcaat cactcaccca tctatacatc catccaccta cccctttctc cctccctcct 1980

tctttccctc cctccctttc tttcctcatt cagactgttt attgagagcc ttgaaggtca 2040

ggcacttgga tggctgctgg ggagatgaag cttgaagatg tgtgaacggg ctgctcctct 2100

tggagacatc atcttttccc tcatcagtga gggttctgga aggggaatgt gggttgggag 2160

gcaggcagga tggagaggtg gctgttgagt tcctgccctt tgggttttgg gtcacagtac 2220

tagactcata gaattgggtt aaaaacttgt attttttttt ctaaggagaa gctggaaagt 2280

gaacaagtaa gatagggcgt ctggctcctt gctgtgtata gaacagatac ccagcaatga 2340

ccataggtgc agagtttatt tatgtagaga ctttttgaat ataagaaaag aaatgacaaa 2400

tatatatgag ttactatata atgaatataa gtataattaa aataaaccaa ctataggtag 2460

tgcagggtga gtttccaata ataaattcta aaccaacttc catcattcat ggaaccagat 2520

attctattca ggaattaata tgcattctcc catacatttc ctattataca catgggaggt 2580

agctactcat attaaatcaa tatcaaatag atgggaacat gaggagagag attagctacc 2640

aaggtaaatt gtcatctctc agtggcagag aaatttgtca accgaggtct tcctagaaat 2700

atgctctgac ctaccataca attcttcctg cctgaggaac tggtaaaaac tctccaagtc 2760

catcctggct tccttctttc cttcccaact tctgctcatc ctgacctgac cttgtcccaa 2820

gctgctctat ggcttttatt gctctttgca gaaatcgctt aaactttttg tttgcagttg 2880

ctccctcctc tgtgtttaac tgtattttaa gggggggttc attttccaat cctctttgct 2940

aaatgtcggc tgctaggtac tcatcccccc tggaatgccc tctctttcca tttttatgac 3000

tcagcaactt tccctgggct tcctgcctct gagctggctg gagctgtgag acaatttcgg 3060

gctttgggca gcacctaaga catacaggat gcgaatccat gttaggatcg tagtgacctc 3120

ttccatataa tcccccattg tactactcct caagagagaa cactctgtct tctcatcagt 3180

gtaatctcaa accctgcacc ctgactgttg tgggtgtagc agacacacaa gaaatatttg 3240

ctgaatgtta agtgaattgg aataaacatg ttgatttaag tgacaaaacc ttctattttt 3300

cataataata aatgtataat gattgtcaaa aaatgaaaaa agttagatgt acaaaatgaa 3360

aaccaatatt acctacagtt tacccccgca accggtagcc ccggggaata gatgtggata 3420

cggatgtgca tctaggtatg gataaaaatg cagggattca tgcacacaca ggtcacgtct 3480

tatctggtgg cctctttggg ataatatgtt caaggctttt cagatgccat tcgcatcatc 3540

gaatacagca tttttatagg atcttacatc ttaaattggc ttaatgtttt gttctccgat 3600

tataaaatct gaaaacagga aaaattaaat aagaatacaa agcaagtctc tgatcctgca 3660

ctgcccaaga ggccctgtta gcgcgttggt atggcgcctt ccagatcttt ttctgcatga 3720

aatcatgggc ttatcacaaa gatgggaact tctttcctca cttcttcttg ccctgttgtg 3780

aacatctgtc tatggccctt ctccaaggcc atctggggat gtggatgtga ggctggcatc 3840

tgctctcggg ctcttttctg tcacgttttg ctgttagctc cctggctaac atccacatgc 3900

actggctctc tggcatctcc atgataaatt cttatgagca ggatttctgg gtcaaaagag 3960

agtgcattcc caggctgtgg agacagggga cacgttcagg agggcctaaa aggctgaacc 4020

gtgtctcagc cttgaggcct ctcagaatcg ccagcttggg aagttaggag gcctgtggtt 4080

gctcatcact ccctcaccaa ctcgttctgc ccagggaagg tcctggggtg ggggcttcag 4140

agccctgtcc tggccacccc cctccctggg gtcctgctga caatcagggc cactgctggc 4200

tggttctgag atgccttctg gtggaccctg ggacccagga tggagctcag ggtgtgggga 4260

ggcagccagg gggcagcatc aggcccaggg gctggactag gagggggtgg ggtggggcgc 4320

aggctctact gcccccacct gtgagctgca caaacatcca gcagcagccc tgaaactgcc 4380

ccatgctcct cctgggccac acctgggcct gtttgtcact catcccatgc ccgggtggcc 4440

atgagctcag tttctcttcc tcttattttt ctccttttgt cactctgagt tctggtttca 4500

gccaacttgg ggttaaattt agcctgggga tttccagggg tggccagctg caggcagggc 4560

caccagagct gggaaagcgc atcccccaac cccatccctt ggcctggccc ccaccctact 4620

cccatctcag gcctccctgg ccaacccctt cacccaggaa tctgctggtt tcctgaatgg 4680

ctggcactcg gggtccctga cccagttcct ccttgaccag agggaaggct gggcccagga 4740

aggcaaaggc atttgcccag gttcacacag tgtcacccgc aggcccaggc aagagcaatg 4800

actgagggta ggtgagggtc cccataggcc aggcacagct taggcatcat ctcaatccgc 4860

aaacctacct tgtggggagg gactcttatc acccccttgg gcagatgagg aaactgaggg 4920

ctacagaaga gaagcagatt gctcaaggtc aagggttgat gagggctgag catggtgggg 4980

ggaccccttc tctgtctgac actgaggtct gggctcccag ggcagatgcc cctttgggaa 5040

gaacagtgct ctgggctccc acaggccttt gcagccaccc tccttccttg catctcccaa 5100

ggcacaggga aggaatggcc atcagggcct gtagccctaa ctctcatcca tgctgctttc 5160

aggcctcttt tctgcacccc tgaaatgggg ctggagctgt ggaatggtgg aggaagtgaa 5220

caatctggtc tctggcagag ggcagtggtc cacacagcca ccctccatac aggtcttctc 5280

catgcaggct cctccacacg ggtcccacca cacagcgccc tccacacggg cccctccaca 5340

caggtcccct ccacacaggc cccctccaca caggcaccct ccaacagatc ccctccacat 5400

gggcccctct acctggcacc ctaccgacag gcccctccat gcgggcccct tcacacatga 5460

tccctccacg cgggcccctc cacgcaggcc cctccacatg ggcccctcca cacatgattt 5520

ctccacacag gcccctccac atgggcccct ccacacatga tccctccaca cgggcccctc 5580

cacacatgat ccctccacac gggcccctcc acacatgggc ccctccacac gtgggcccct 5640

ccacatgcca ccctccatac aggtcccctc cacatgggcc cctctacatg ggcccctcca 5700

cacaggggcg cctatgtgag ctcgggcaga tccctttctc tcctggtctg tttctccatt 5760

tgtcagatgg gaggccctgg gctggaagtc aggacaccac agagctggct ctggcttttt 5820

gggatctaac tgctctgggc ctcagttttc ctacctgtga aatgggacca tgattctggc 5880

tgtggcctca cagaatccct gagtggctca ggtggataac atgtgtccac gtgttttata 5940

aatggtacgg gacgataagg ccactcaata gcgtctcaac acaggatgca gctacctgta 6000

gagttgcaga tgtggtcaga cctgaggttt agggcctcta ctctcggcct tgctctctct 6060

gggccctgcc cctgcccctc tgcagagctc tggggcttgg ggcactctgg tccactggcc 6120

tggaccagca tgaatgttca tggccttggt ccaccgtcca acccagagaa ctgtgcaccc 6180

tgcagggccc tgccttgggc ctggctccca gcatcctctg ctcattccct ctcctccacc 6240

gggtggtcac tacctgtgtc ctatgcaggg cccagacctc ttttctccct tgttccctgc 6300

ccccctggaa gcctggtcca tcttggagcc catccatgtg gcacagactc caaggacttc 6360

ctgggtatta agtagggtgg caggccaggt gacgtggggg acagagtatg gggcctttga 6420

ggtgagcttt cagggtgctt ccatgagggc ccttgtgcac tactgtggcc ctgtgggaag 6480

aagggatgct gtggtcactc ctataccttt gctcacactc acccctaaca cacccgtcct 6540

ctctctgccc ttcagtcctt gatccagctc ctaggcctgt ttgaggttgc tgctgccacc 6600

aggctacctt tgatgaccac atggaatccc cagtatctcc tgccccttgt cttcttcaag 6660

caagggggtc tcatctgcaa tctgagctct gggctgggcg gggacaagta cccccccttg 6720

cttgctttac ttcaggaact ggcctgggtg gctggcatcc tgtgcatgtt gaggggacgg 6780

tggtggggga tgccagtgga tgagctgatt gaagatggtg ctgggtctag ggccaccctg 6840

ttcagcctcg tgggtgggcc tgtggggagg aaggcagagg tgggggcact catccctggc 6900

ctgcccaccc tacagttcgc atcttatgtg tcaggtgaca tgtgcaatgg ctacgtgtgg 6960

cgggcctggc tcctggcttt cctgggccaa gggagctcca ggctggtgtc ctctaagaat 7020

gagctggtga agtcccacag gtggatggga tttgccgtct gcaggggtga acccaggccc 7080

tgaggctggg tacttggtct cctcccatac tctcatcaga acctggagcc ctggaatatc 7140

ctgggaacgt ttcctccaca tggagaaaga cagggaggtc tgtaacgggg gcagtgggta 7200

ggatctggtc atcggacagg gggcattaag cctcaccaca gtggcctgtt cctcacacct 7260

gtcaacctct ctttgacctc agcatctaga cccagcatgg ggaagactgg acagggaatg 7320

gggcacccca aggccacagc cccctcctcc catgagaagg cacactccag gtctcacctg 7380

ggggtgggct ggggcagtcc tgccttgcct gggacaccat gtgtcaagag ccctgcacag 7440

gccaggcctc ggggcttgtg gaaggatgag tgtgtgtgtg tgtgtgtgtg tgcgcgcgtg 7500

tgtgtgtgtg tgcatgttca gatatatgga tggacagaca cacaaataag tgactgacca 7560

gtctgaggtc ctgcccactg ggtagggacc ccggaagtcc tcttggcagc tgtgcttcca 7620

ggagcttcgt tactgcctca ggctctgggc tcccagggaa gctcttggaa gggttcttgt 7680

ttcccttccc atccccttac ttggcccagc tcccttgaag ggtccaagac ctgggggcag 7740

caggtggagt gtgatcagca gggtcccatg tccagctctg gccagccctg cccaccccag 7800

gcccctctga gctccaagtg tctggctgtg ccccccaatt gggcacagct ctgaacctgg 7860

catttagaac ggttcctggg gagctgtgag gagggggtag ggcctgcatg aatagaagct 7920

tctggagcag aaggtgctac ttctgatcat tgattgacca tgatctccag accctgtcta 7980

actcaagcct gggcacgggg cttgggtgtc ttggtgcggg gacacaggac cattctccag 8040

ctgggctggt ggaagctttg gaaactgaat cgcccccctg cctcccaccc tggggagaag 8100

gaggcacctg ctgccagggc tgaggcttgg tggggtttcc cagctatttc gggcctcagg 8160

cactcttggt ttcaccaggc cagctgcact ggggtgaggg ttccagggcc ggtggggagc 8220

ccagcatggg cagggggctg gcctgaggct gtctggggct ggaacagccc aaggatgaaa 8280

acaaccaacg gcagagacaa acaaaacatc tattttccat ctgattgtcc ctccccccac 8340

ccagagagtc agtttccatt attatcaacc cagctgcaca gaggggaaac tgaggctgag 8400

ctcgagccac taactccagc cacaggggcc tggcgctagt cagggctcct cagtcgctgg 8460

ctgcctccag cccctgggac gtcgcccccc gtgagacccc agccaggtgc tccctacccc 8520

ccatcctctg ccctatttct ccccagcaca aatcccgcct cctacatgac attgttcctt 8580

gtctgttgac ttgatttccc tctcccgcaa acatgagctg tctgaacctg gctgtgcctg 8640

ccttaaccct gctgggtttc caggagcagc aggtcctcag aagagagctg aacatgtgac 8700

ttcgctctgg gcagcccctg agaagggggc ttacttacct ggccggtctg gggtctcacc 8760

ttcttcagct actctgccca gcagcccctg gggaccagca ccccttcctt ctgggagctg 8820

ctctctgctg cccagacctt tgcaaacttt ctgtcccctg tgggagccca ggccaagtct 8880

gaccaccact tcccatcctc ctccccagac aggtcaggag aggatgtgtc taggggtctg 8940

agtgggaggc tcccctttgg cctgaaactg tgaagatgag gcccctttct cctctatcat 9000

ttccatagtc agaaagtccc aacatgtccc tgcacaggct gggaactggg tgggcaggga 9060

gggcccatcc tagtgacacc tgtgaggggt ccaggctttg gtgcctggca cggcaactgg 9120

cacattggat gctcagtgaa catcagacct gatgtgagaa gcaggaggga cccgtggcca 9180

gatcaagggt gttggtgttg ctataggaag agagagtcac catgaaggat ggcctgtatg 9240

gaaagaggca cactgcgggt gccctggtgg gtcgtcctgg ctgtgggcta ctctttcctg 9300

gagctgacac gggggctctg tgttgccagc cacagaagtc acccagcccc tccctggctc 9360

ctgtcccaaa gccaggaggc attctccctg ctgcctgtgg tctccccagg aactagggct 9420

gcccagagga gctgtccctt gccggtctct ggacgtggtg tgctgttgct gagtgggaga 9480

tggtattagg gccacccctg gttgtgttgt gagctgggcc tgagtgagct catttctagg 9540

ggctccaggc aggggaccag gaatgtcatg gtgactgcct ggatgtggcc cctagtgagg 9600

ctctcctggc ccctggcccc cattgtaatg tccagcactg cttaccatag tgctaggtgg 9660

ctgggcttgg cttcctcccc ctcattatta gccattatga agaacgtcat tcagtcattt 9720

agcaaacagt tactctgagc ttgggcaacc aaaatttgtt gtttaaactg ggatgttcct 9780

gggagtggaa ggggcactat taataacttc acgagtctct ctgatagatg tgggtggggg 9840

tcctggaggc agcccaggcc tgggtctctg ggcagagccc accattactg ctacaatggg 9900

ccaggcacgg ccaggcctgg gtcccgagct gaggctgggt acacataggc acctgggaaa 9960

ctgcagctga ttggtggcca gtggtctggg gacctggatg gggctaaggg tcaccagggg 10020

ctggacactg ctctagcatc cacctcagct tccagagtca gtggcacaaa ctgggaatgt 10080

cctggaaaat gcaggatgca ggtcatcccc ctcccctcag accttgccca gcccacgtgt 10140

ttcctgggga gaccaccctt gtgaaattac tggccaaaca gggtcaggtt tgatgtggtc 10200

ccaagtgacc aagtgacaaa gactgaccct gatcctgacc ctactgcccc tgctgatgtc 10260

actgtgttga catttccgtg gcccatctgc aatcctcaag atgaccatgt aggagagcag 10320

tttccccaga tcaccccaga aagagggagt cctcacgctt gggttgttac aagaggtacg 10380

ggtgcctttt gaggtgtttg cataactaag cattttggga aaagacctag tcttcccggc 10440

ctccgttgat gccagctagg gatgcttctg aatccccgag ggaactggag catctcacgt 10500

gtggcgcagg ggatcctggg cagatttggg gtcatttttc cttcacgtct cattcagcac 10560

agggaagagg accagtgtca agtcatgaca cacgactctt tcagggccgt agaagctggt 10620

ccttaggtca gggatggagc ccagggtgtg gggtcaacat caaggactcg tgtcctgcag 10680

gcggggtggg tggggaatcc agaacctggc cctggtgtga cagagactct ggagaagagg 10740

atgacatgtc gggagggagg agcaacatcc tcagcattgt gctgtgtcat tagcagtggg 10800

cattgcgagt ggatggtgat tatttcctct atgtccccaa aattcgcatg ttctagttgg 10860

ctggaaactt gattttggac ctttgtgttg aggaatgggg gcttgcctgg gatctggtgg 10920

cctcgttatg gggcatctga gctggtttgc ttgttcccag gtgagtgtcg atgtccccct 10980

tcttgcccct ggattcagtg aatatggggg ttgaagggta atggtgcctt tgtccaggcc 11040

tggctgggga cactctcttg gtggctttgc aggccccaca ggaggctggg cctgatggga 11100

gagtcaccca ctcagttcat agtagctccc agtcctttcc tcccgcaggg ccctgagcag 11160

ccacccccct ctcctccagc agggcccctg caccccctcc caggctattc tgggctctgg 11220

ctttgctgta gaggaagcca cagggcccct ccctgatctg actcagaaca cctccctcag 11280

cccaggctgc agctaagcag ggccagggcc aggacgagtt cctctcaccc catcccccac 11340

tgtgacactc ttccctatac cttgaggggt tcccccagca cagaccaacc ctccttcggt 11400

gtcatgaggc catcactcgt gaggggctgg ccccggtctg agctagacac agtggcatgt 11460

gaagacaggc gtgggctaag gcccaactcg gctactcctg aactgggtgg agcctgggca 11520

gcgctggtgc atccatcatg gttcacgtcc cctctgtgag gtgggcaatg gcagctttag 11580

gcctcagggt cactgtgagg gtcagtgagg gactgtgatg ttcccactgt ggcccaagga 11640

tcacagctgg ggctctctca gatggttctt tcagttaaat cttttcctga gtgtggggag 11700

atgctcctgc tgccccagac actccctggg cagagtgcca accttggaga cttgacaggg 11760

tgggggcagt ggtgtctcca tgtcgttctc caagcttgtg agctcctcta gggcaggtgg 11820

tcctgggctg catacaccag cccctggccc acagctgaga gtttgttgaa tgaatgaatg 11880

agtgagtctg aacaaatgaa tggatgaatg aatgaatgaa aagcccttag ggccgaggag 11940

tcatgaaaag aggcgggcat gggtcagccg gtaacctact gtgtgaccct aggtgggtca 12000

tcacccctcc catcggagcc tcagtcctct catctgtgaa atgggcacat gaacacctgt 12060

cctggctctg ctttgtagac tcagagggca cccccatggc tgccctcctg cctgcctctt 12120

tcacccactg agtcaacaga gaatgaggag cccaggtctg cccatcccac agagccacaa 12180

gggggttcat gcactcagca ccatctcttg gccttgccca ggcacggcct caggatggaa 12240

gggtggccta ggtcaggggc ctgggcaacc actgtgctgc cctgaacacc tgcaggacct 12300

gggcaggctc agcctcctct gcacgcctta gggaaggggc tggctctggg ttgaccttgg 12360

atcagagtcc cagtgcccac catctgtgca gccctgggca gctgtgggac tcgggcaggc 12420

tgctggtctc cgaatctccg tctcctcatc tgtaaggtgt gcgggacatg gacacgtgcc 12480

tcctggggtg gtttgggatc ccatgacata gagaggacag acgccagtgc agggtttccc 12540

tggaagtggc cctgggccag gtggcaggtg aggtagcatc ttcagcagag ccgggagcag 12600

ttccactgtc gagcagtggc catcgagggg ctcagtgata cccagcccag cacctggctg 12660

aacatcggag gacgtgaggg gtgcagcccc tgcttccagg ctcccgtggt gtcccagatc 12720

ggtgctgggc agggcggcac caggagagcg tggtcagcag ccctgggcag ccatcaagcc 12780

tgaggcacca ctgggctgag ggctggggcc ggggaaggag caaggaaccc ttttacattc 12840

cggtagttca tgaatcacac agtgaaagaa agagtcacca agggtgctag ctgccggtgg 12900

ccctgatggg gcagcagcag aacaaagtgg gaggggacca tagaagagag gaccttggcg 12960

gggggtgggg gtgggggtgg gtgggtgagg ggcttcaaga tgacgagaat gatgtgagtt 13020

gccaagccca tcctgaggtc aacagaagga ggacagtgag gggaccattg gattgagcca 13080

cgtgcaggcc actggggctg tgatggggtg gtggtgagac tgctgggctt gaggcaggaa 13140

gggaggggac cgtggaggcc aggagccctt gattaccact tgtaaaaggc aggagagagg 13200

gaggggcagc cagaggaagg cagcattcac attttattac ttcttcctgg caggaccaca 13260

tttgtgctcc taaggtgacc atcaggaggg tctctggccc ttcatactga gatgttgagg 13320

cagagagact gtggtccagg ttctctccag gaagagccca ccattactgc tgcagtgggc 13380

caggcaggac taggtctggg tcccaagccc aggctgggca catatgggtg cttgggaaac 13440

catagttgat tggtggccag tggcctagta cctggatggg gccggggtca ccatgggctg 13500

gtcactgctc cagccccagc atcaacccca gctcccagag tcagtgggga aggggactgg 13560

agtggggtcc gattacagtg accacaggga aatgggggcc acagggaaat gggtgcgggg 13620

agcaggggag attcgggagg tgggtggaag aggaccacgt gctcccatcc tgacagccca 13680

gagcctgcgt gtgcctgggg cggctgctct ctcaaagggc atgcgtctgt cttggggact 13740

gagggtctct gaagtcaggt cgcctcttgc tcgaaagccc cctgccacct tgctcttgac 13800

tttagcacct cctcacaggg cagcacgtgt tatgttatcc gggaggaaat ggaaaccctg 13860

ggaggttctc tggtggggga agggaggagc ggaggtagac taagaggggg tcagcagggg 13920

gagaaggggg aaccagccca gagttctcca gggacgggtg cgtgttgtcc tagcctgtgc 13980

cctgggagtg acaccaggtg agttgagatt tcagtaccca gcatttttgt gactgaagta 14040

aagaaaattc tgatttggcc cttgcctgct tgtacaagtt aactgcttgc tttttgccca 14100

gcatcccttg ttcacacaat gtgatcataa gctgcagatg tcctgtttct ttgtcaagaa 14160

ggaggagata acttcagagt cacaaaagta ttttgcagga gtgaatgcct ttagggacgt 14220

agagaccagc cgctgtcgcc cgcaagtcta gttgcttaag atgttacctg agactgaata 14280

aacacagact tttcccctga aatgccacct ccctcactcc ttagccacat aaaaactccc 14340

tgcttcatcc ttttgttaac atggagtgga ggaatttgct ttcttgtctt cttactttag 14400

ccaaatgaaa taaatctttt tctatctcca agcacttgtg tcagtttttg acctccgctg 14460

catatcaggt acacgaacct gaatctgggg ttctacaaag agtctcacat gtcaccactc 14520

agcccccctc gaacacctac tgcgctcggt gctgggcccc gagaggaccg gggttcctct 14580

ttcccacagc cccagaccca gagccccatg tgctcttccc ttgggagagc agtccctgac 14640

tgaagatgtc aggaattttt ggatgttgag ggagcaactg ggaccaggaa tggaggggat 14700

tgggcttggg gagccacctg ccagggactg actctcctgg aggtggggga caccgtaagc 14760

cctggatcca gaaccaggac tgagatgggg tctctaggaa aaggggactc tcatggtccc 14820

tccctccgag tgccctgacc ctgatatttg acccggagga aggagaaggg gcatgaggaa 14880

tctggctact ccaacctgag gccccgtccc cttccatccc ttgttggtcc aataattctg 14940

tcacttgaat cccgggtccc gggatggggg cctgggtcca gtgggctttg agctctccag 15000

ggaggaagct ggatgaagtg ggtgggcgtg gccaggtggg ctgagggcag ccaggcacct 15060

ggggtccaga gctgagatct gcagttgggt ggaccaggtt tgagtcccta ctgtgacctg 15120

tggcaggtca ctcaggctct ctgggctgtg atgtcaccat cctagagtga ggatcacaat 15180

aggatcttcc tccaagggtg atttggcctg gagggcattt agtctcaggg ggcctgctgg 15240

aagacagtgc cctagagctg tggctaaact cccagtgcct tcctggaacc ccaggaggag 15300

aaggggccac aagagaccag ggatagggag ggcttgggct gggctgggac ggccccccag 15360

gagtggctca gcctggactg ggcagggcat ggagaaaggg agaggaactc gggaaagggg 15420

ctgtggtcgg ctaaattagg ccccgcgggg acagtcgtgt cccaatcctg gacctgtcta 15480

tgtgtcatgt gtggccacag ggactttgca gatgtcatgt caattaggag cctgaggtgg 15540

gaaatgctcc tgggttatcc aggtgggtcc aagctggggt ctttgtagaa gaaagaggga 15600

ggcaggaggg tcagagtgag aggacaagat gtggggatgg agggagacgg agagagagag 15660

aaggaagaga gggagagaag aaggtaggga gggagagaag agcaagagag agatttgaag 15720

atgctgagct caggctctga agttggagga aggggccagg agctgaggtt gtcctctaga 15780

agctggaaaa gtcctggaaa cgaattctcc ccagagtccc cagaaggagc cggccctgct 15840

gggaccctga ttttagcccc caaggactca ttttggactt ctgacctcca gacctgggag 15900

agagtcagtg tgtgatgttt taaggcacta aattaggggt catttgtgat aacagccagc 15960

agccacagga aagtcaaaca ggagccgtgg gagagactcc ccaggatgcc acaagggccc 16020

ctcccaggcc ccacccacag taacccaggc ctgacccctg caggtgcccc cactctacag 16080

aggaggggcc atgcaggagg ggctgggtgg ggctgggtcg tgttctcttc ccccacccct 16140

tcctctcaga gagcagttgg agccccagac tcagggaacc tcacagagaa agtcctgatg 16200

agggacctgg ttttccagtg aaggcccggg atccaatgga gaagtgttcc caaaaatgat 16260

gttcccaaag acggccagtg actttctgtt cacgcacttg agagaatcac catgtctaag 16320

ggctgccaga tagttacctg tgcgtgtgtc aaggttcaca tcccattact actgaggcct 16380

tggatagaac atgttcccac ttttagttgt gacgagatga ctggccccag cattgagaac 16440

accaccttcc cttcagcaaa gggtttgttt aaatttacag aagtggtgaa gtgcttatta 16500

ccctccttgc actggttcct agggagctca gagcctatct tttccaagtg tcccgagcct 16560

cctgtaggcc tttcgtgtga gtctccttcc tggcttcctc ccattcctct taagtgtctg 16620

aacctgctgt caggtgagga atcgaggtgg gtccctcatt gaacagcttg agaagtcagt 16680

ggtgctggag tgtcctggac agtctggctc tcagggatga cagggagggg gtgagggtgg 16740

gtgggagctg agctgggcac agagggagga gggcattccg ggggcaaggg ccaggggcaa 16800

agaggctcca ggcttggagc agggttttca ggctgggaaa agagatggtg ggtgaagggt 16860

gtctctgtta gggcagagat agagggtgta actggagtcc cagccagaga caactccatc 16920

caggcaggag gcagggaggt ggggggctat gctttgagga gaagggggag gaatgaaagg 16980

gcagagggac tggtgttcag tgatcagact tctcctatcc gggcctcttt caccaggaga 17040

caccccggga ggctatgagc ctctccctca gggccacttt ctcactgggt gccaggattc 17100

ccccacccaa cactgagaac ccaggggagg cctcaaatta tgtgtgagat aagaaaagaa 17160

gagagggagc aggatttctc gttgacgatg aggatattta ttgagaaggg ctggatgact 17220

tgggatgggg gcagagaccc ctcccctggg aacctgcagc tccaggcccc tgtgggtggg 17280

gtgagggtcg ggggcctaag agcattctgc aggggccact gtcttctcca cggtgctccc 17340

ttcatgcgtg acccggcagc tgtagcttct gtgggacttc cactgctcgg gcgtcaggct 17400

caggtagctg ctggccgcat acttgttgtt gctttgtttg gagggtttgg tggtctccac 17460

tcccaccttg acggggctgc catctgcctt ccaggccact gtcacggctc ccgggttgaa 17520

gtcacttacg agacacacca gtgtggcctt gttggcttga agctcctcag aggagggtgg 17580

gaacagagtg accgaggggg cagccttggg ctgacctgcg gggtggatga ggggcagggg 17640

gtcagagtcc tggtgtccac ctggggagcc cctgacctca gtcttgaagt gggtatggga 17700

ctaaggccca gggcagggac ctcaggccgg acccttgctg gactgaggtc aggtcagcgg 17760

tgagtggaga cacttgctgt cagatgggag ttgcccccac tcttgggggt cacagacccc 17820

ctcttggatc tgtctctttt tcatattttc catcacctat agacccccac cggtgctggc 17880

tccacactgg atgggatgga gtccaggacc acatctcacc ctgggtattt gtctatcttg 17940

ggggtgtctg tctgaacacc tcctaccagt tccccttgtt ctcatctctt catgcagggc 18000

agctgttggt cctgggttca ggctcttctc tctcacaggg agtgggtttc ttacccacac 18060

atctccctgg tgccatcatt gcagccccca ccccacctcc ctgcagagac ccctctgggc 18120

tgcctggtga gacagggtgt cagtcctggg ggctggattc cagtagatca aaggcctcca 18180

tttgaggcat ttttttttta ctccctgagc tgaacgtgga gctttgtctg ttcgaaagag 18240

tctccatgtc gtgagaagtt atctggggga tctgactctc cagtgggttt tccctgacaa 18300

acaggcaccc gtcccagtca ccgtccctac cagaagtccc tgtcctgggc atctgctctg 18360

aggtcttcct ccagtggagg aattggtaag catccacgtt tagggcctga gttttctgag 18420

acccaccacc ctgtctgtgg tgtcctgcct gtcttggggg atggcgtgtg acccagcgct 18480

gtggggccac cttctcacgg aatattcttc ctctatcttt gtcctcctgc catttcctcc 18540

cccatctctc caagtcctca gtcaaaggcc tcctgtcact gtgtttaatg cagccccgta 18600

actgggaccc acagaggaca gaatcacaga agagggcagg agcccaggca aggaggggca 18660

aagattccag acctccagac aggcttagac cttagccttc gacagacagg aaagggaaaa 18720

actccccgtg tttaacttgg gatctcaaag aggagagaag cggggagact taccgaggac 18780

ggtcagctgg gtgcctcctc cgaacacagc acacagtgac acagccccac acacaaaccc 18840

ctcctgacac acccgacccg ttccccgctg cagcagcttg ggcgcccagg tccagtggcc 18900

ctgtcatgaa gcaggtctgg ccgataggag ggtgaggggg atggcccctc catgcatgga 18960

tgcacacttt actggaagtc agggatccct tcacccccag agggagaggg gagagggagg 19020

aggcatgtct gcacatcaca tctgggtccc ctgctactct ctgtcctgga ggaagccatg 19080

tgtcccggct cagctccttt ggagactgtc accatggaag aagtgagaat cctgtttatg 19140

ccacagggaa ggtggagctg gtcccctatg gagatccgag ttccctggga gggtcctgga 19200

tgcccagcac agcccctagt gggatgccca tgtggtaatg attctaaata gcacaggaga 19260

gtgtgtgtgt atgtatttgc atgcaggcat gtttgtttgt gaatacctgt gtgcctatga 19320

ggtgttcatg tgtgtttatg tttgtgaaca tgtatgtatg tgtgtttgtc tcatagccca 19380

ggccccccat cctgtgctac ctcctgggac aatcatcaac ccggttctta gatcccacat 19440

acagctctta aaaagaccaa ttggtttaca ggcacgtcta tggcaccact ctggaatctc 19500

atgattgact ttgacctccc cagccccact gaccatccgt ggataactgg acattaggta 19560

cttgaggtgt ggtcagtttt gggctggtta cttaggggtg agagagagta gagttgaccc 19620

ttatccccca ggggacttga gtataactgg gacactggtg acacatgtgg gaccaggggt 19680

acccagctgg cctgtggggt gagtgacctg ggtccagagg gtcagggtcc agtctgtgcc 19740

tgcttcagag atagatgtgg gggtagaggg cagggtcctg ggagtcagga acagggtggg 19800

cccctaagga gtaggggaga ccaagggccc atggactgga gaggctcctt ccgcctcaga 19860

aggagggacc ccttgagaga gggaggctgt ccaggggtga ggcctgatga tccgtggagg 19920

gtactcgttc ttctgggcca tggggaacca aggagcggtc gccccaggcc gctggaggtg 19980

gacatgtcca cacaaacaca gcacgtatgg gaaaggggca gtcctggggc ctgctgccca 20040

aggctctgtt ctggaatatt ctctgggcct ttctaggtga tggtggaact acccctcaaa 20100

aacaatccag acttcattcc cacaaggcta aaagaatttt gttcatcttg caggttggac 20160

caatttaggg tagaaaatgt ataaacagaa gcaaaacaca aaacccaggg tttctggttg 20220

actgaggata tttattgagg gtttattgag tgtaggaaga agggctggat gacttgggat 20280

ggggagagag accccctccc ctgggaacct gcagctccag gcccctgtgg gtggggtgag 20340

ggtcgggggc ctaagagcat tctgcagggg ccactgtctt ctccacggtg ctcccttcat 20400

gcgtgacctg gcaactgtag cttctgtggg acttccactg ctcaggcgtc aggctcaggt 20460

agctagctgc tggccgcgta cttgttgttg ctctgtttgg agggtgtggt ggtctccact 20520

cccgtgttga cggggctgcc atctgccttc caggccactt tcacagctcc cgggtagaag 20580

tcactgatca ggcacaccag tgtggccttg ttggcttgaa gctcctcaga ggagggcggg 20640

aacagagtga ccgatggggc agccttgggc tgacctgcgg ggtggatgag gggcaggggg 20700

tcagagtcct ggtgtccacc tggggagccc ctggcctcag ttttcaggtg ggtgaaggac 20760

taaggcccag gatagggacc tcaggctggg cccttgctgt actgaggtca acttggcggt 20820

ggcggggatg cttacctcct gatgggaaac tcctcagcat ctgaggactg aggagaatca 20880

ggcctcttct gctccagcct cctctccaga ttctcccctc cttatcagcc agctctcacc 20940

tcacctatgc tccccgacgc tggctgggtt cagccctggg ccctcgtctt tccttggggg 21000

acgtctctgt ctgaacatcc cctgccaggt ccctcctctg ggtctgttca cgagggcatc 21060

tgtgtcctgt ttttaggttc tcctcccttt tgacaagtga atttcttcct cacatatccc 21120

ccagcaccat cattgcagtc tctgccccac tttcccacag aggccccttg gtggggaggg 21180

gatgtcagtc ctgggagccg ggtcctggga gtccaggaga tggagggctt gcatgtgagg 21240

tatattttct gttctgagcc tggcgctgtg tctgctctat ggggtctccc tgtcgttggg 21300

agatctggct gtcctaggac aaatgtctgt cctgacaggt ccctgtcacc tgcgcccacc 21360

aggagtgcct ccctcagcat ctgctttggg ggtctctctt tttgggtggg atggggaggc 21420

tctaaccttg gactgagtgt cctgagaacc accctgtcca tggcagcctg cctgtcccgg 21480

gggaggagga gggacccagc accctggcgc cacctcacag gacattcttc ctccatcctt 21540

gtcccccttc atttccccaa gtccccagtc aaaggcctcc cttcttggtg tttaaagcag 21600

ccccatagtt gggacccaga accctgggaa gctgaacttt ttccagcatc cttcccctgt 21660

cccatggccc tggtgcccca gcctgagccc ctgctctgct ggtctttggg gactggagac 21720

tctgtgcgag ggagcagaac atcccagagc ccaggggaga atgccccctg agtgacagcg 21780

gcaggttcag gcctgagcag ggagcattaa gaggtcactg ggatcttccc atcatgcccc 21840

accaggtgcc taagaaggga caggctggag gaatttgggg ccaggaaaaa aggcaatggt 21900

ggcagacccc agagagattg agacctctac cctagagaga acagaaagga cagaaaagtc 21960

tccccagatc tcaccctaga cccaaaagaa tagaaaaggg gactcaccga ggacggtcac 22020

cttggtgcca ctgccgaaca cattacactg tgatataacc ctgcacacaa accctcctga 22080

tacacccgac ccaggccacc cccccccaac cccctgcagc tgctcacaag gcccagggtc 22140

cagccccctt ttctcatggg tgggtgagcc ataggtgggg tgaagggggt caccttctac 22200

tgacctgaat atcacttcct tgggggctta ggggttcatc cacctcagtc atgggaaggg 22260

ttgacaggtc tgtgtgtgga attgtctccc aggtaattgc atttctgagg agttgggtgg 22320

cctggatcag ctggtgtcag gaatgccagg gaggaagggg aggtgagcgg ctgggtgacg 22380

ccactgacat ggtgcagctg ggacctcttt gagaagttgg gaatggtggg gagggtcctt 22440

gatgcccccc acatcctcag cactggaggt tatgatggga tgggcttgga atgggaattg 22500

ggggttgtgg gaagaatgac gttgaccaca gggacacaga ccccactgca aggtctgagc 22560

tccagcatcc ttggcccagc tgtcctgaag gagcagctca cagatcactg cccaagtccg 22620

cctcctgaac ccatggggac agcttcctac ctataggcca tgtgtgctgg attagagctg 22680

caatggccca gaggagagct tctcaaggga ggtcataggc ctcactggca gtgtggtttc 22740

ctccaggagc tctacgtgtg ggggatctct gctgtccact tcagctcccc cggcttacac 22800

acacacccac atgtgtgtgc tctgcagtcc cctacatgca cgggtgaccc acacagcctg 22860

tgctctggag tgtcaccggc acggggggca cccacagtgg atgggcatgt gcagtgtggg 22920

gcctggacac acacagcctg gacctgcatc ctggtctggg cccatcacct gaagagctca 22980

cacctgggac acactcctgg caccatgcag tccccaacag tattccccac agcaagcgcc 23040

tcccctgact gctgagatgt ggcctgctgc tccctgatca gaatcccgtc cacatcttct 23100

cctgtctccc tcttggcctg gacagaaaac cctgccccac cctcccatat ccaggggcga 23160

ccccctagtc ccaaagcctg gagacagcag gtctcagctt tgctttgctc atgtcctgac 23220

ctgggcctga ctgcagcccc ttctaccttc ctgatcctcc cagaaggatg cccagcatgg 23280

acatgtgtgt ttatgtgcat tgtgtgtgtg tacacgtgtg tgtgtattat agctcagatc 23340

ccatgctgag ttcttcagaa gaagaatcag cgtcacccaa gctcctggca taaccccaga 23400

atctgacttt gaactcacca gtcaattgac aacttgtgca cagcaggaca ttaggcattc 23460

ataatggtgt cagtgaggag cagagccgac ccttgtcccc taggaatgtt gaatacagct 23520

ggggcatccg gtgacacatg tgggatgaga ggcacccaga gggggatgtg gggtgagtga 23580

ccagggtcca gggggtaagc atcccatcag tgattattat agggagggcc tgtgggggct 23640

gagggcaggg tccctgggat gacgggaaca gagtgtgccc atggagaagg ggagatgctc 23700

taggctttga tgaggggaaa tacaggtggg gtgaccacat ggccttggag ggggatggac 23760

aaacaccttg agagagggat ggcatctgga cagtgagggc cgaggacaac aaagaaggga 23820

gctgagatgt tctcccgggg ccatagggat gtaaggaggg tgtgtccccg ggcagccaga 23880

ggtggacgtg ctcacagggc agcaggaagc acaacagaga cgtgggtaag aatgaagatg 23940

tctggttcct tctgaagact atttccttgt tctggaacat tctatgggga taaggtgatg 24000

gaaaaagtat ggctcagaga aacacgaaga tggtcccagg ctgcattgtc acaagtccaa 24060

aggattctgt tgatctttgg agataaaaaa atttaggatg gaaagtggat aaacaaaaat 24120

gagcagacag catgttttct gattgacaag gaggacattt attgagggtt tattgggtgt 24180

cggcagaagg actggatgac ttggggtggg agggaacacc tctgccctgg gatcctgcag 24240

ctccaggccc ctgtgggtgg ggtgagggtc gggggcctaa gaacattctg catgggccat 24300

tgtcttctcc atggtacttt ctttgtgcgt gacctggcag ctgtagcttc tgtgggactt 24360

ccactgctcg ggcgtcaggc tcaggtagct gctggccgca tagttgttgt tgctctgtgt 24420

ggagggtgtg gttgtctcta tgcccttggt gactagggtg ccattttcct tctatggctc 24480

ctgggtagaa gtcatttatg agacacacca gcatggcctt gttggcttgg agctcctcag 24540

aggagggcgg gaacagagtg accaaggggg tggccttgag cggacctgca gaatggatga 24600

ggggcaggag gtcagagtcc tggtgtccac ctggggagcc cctgacctca gtgcatgagg 24660

agtgttcagg ccccccagta ccagtgcagg acctctcaga tgtggtccac actgtctcag 24720

aagggtgtga catcatgttc ctaaaagaac tctgtcagtc agaaaacctc tgagcttggc 24780

ccctgcccat cgcctgtttg gaaatcttcc aaggacttgg gcttttgtat agacagtgga 24840

ccttcaattg cctggcgctg cctcaccctg tcatcctgac gtgatccggc tctggctgtg 24900

ttctctgagc tctgtgagcc tcttgttaga ttcactctgg aaacttttct cacctgacac 24960

agctgctctg ggttggggga gagtgggtct ctcacgtaag agctcttgtt tgtacccaca 25020

tccgtctctc ctggatcttc tgcagtggag tcagggcctt tgctcaccca agtcccactg 25080

tctgtctgtc ctgggctctt tgtctgtgga gtctgtgtta gagtctctct gtgtttaggg 25140

ccaatctaca gtttttctgc cttgggatat gcctgtcggt gaactcaggg gttcctgctt 25200

ctaggggtgc ctcccacaga gacccatcag ccacctccct gaatgccatg gcctttcgtg 25260

acacttcacc tggctccttc atgtccagag ccttcccgtc cctcctgaca gggcttcatg 25320

ggtcccaggc cagctcaggg ctctctccct gggagcccac agaaacctgg agaggcccct 25380

gcacctccta ccttgttccc agcctggatc cagagtccca cagtccaggg gactgggccc 25440

tgagactctg agttttcatt cccctatctt ggctctgagg gtttcaggtc cttctaaagt 25500

ccaccaggtg gtcactcctt cgagcccctt gttccctctc cctcatgaga tggggcctca 25560

gtttcccttg cacaaagttc gttagacaga agcccaggtg ggggaggaca gaactgagca 25620

agttttaggg aagcagcctg gctctggcag acagacagac agatgagtct cagtaggtca 25680

tccagtctag tgccctacat ctgaggtcag aggtgaaggg gtcttggtga cccactgagg 25740

taagggtcta aacagggagg ggagaggtcc ttgaccaggg cctaggctcc agagagaaag 25800

catcaccaca ggtcaacaca agatacagaa aagaacaaaa gcagcagaaa atttcccagt 25860

accttgggga gaccagggaa ggagggggaa aagactcatc taggacggtc agctcggtcc 25920

cctcaccaaa cacccagtgc tgtgacacag gctcatacaa aaaccctctc tggacgcctc 25980

tacccccaac ccccgctgca gctgtgatgg aggaagctga ggccacagcc cctggtggca 26040

tctgctcatg agggagttca cacggctgcc acactcaccc ccacagagag aaggggacag 26100

ctgacatcct gtgaccagca ggatccctag aaatcctggt tcctggagga cagtggattt 26160

ggttttccca ccaaccatca tgcagccatg gggatgagtt tattggaggg cactgtgtca 26220

ctaggagtga actcccatgg tcagggataa atggtctccc tggtggagct gactcgacac 26280

tgactgcttg gtgacgccct ggggtgatgc agagagtgcc ctctccctcc aggaagaagc 26340

tggttaccct agagtgcggt gtcttgggaa gcttcccatg aacctatctg tgactccagg 26400

agcctcagcc cagccttccc tgggcttgtc ctagttggga tgggcctggc cagtgaggcc 26460

tggctggtcc ccactggaaa tgccagcccc atcctccagg gtaaaatgga aggatctcct 26520

gcacccctca acctggtctc attttggtgg gtgaggtggg gtagtggctg ccactgcctt 26580

attcaaatgt gagggacata aaggagctcc cgagtgtggt ctccaaatat caccttctgc 26640

taagggaggt gaggtcagtg cgggatccca ggcctggggc aggaaaggga catgatgaga 26700

cagaacctct catcctccga gaaatcagga gccatggaag ctcctggggc aatgggagaa 26760

aatcagatgc cacctgagga ggcgcccagt gggtaaatgt ttatgttttg tagccgcatt 26820

gtcattgcaa ttcagtatat ctgcagtgga gcccacatgt cagatgtcct cacacgctag 26880

agctcactcc ggtactacat tgtgctgtgc gccctggagg atggctgggc accagatcat 26940

cagttatcaa agtggtgaac aaaagggaag aatgtacatt ttttcttctt cccaaagcat 27000

gaagcacact cctgagcgca cggtgcagct gctgtgccct cccttctgga agccctcatg 27060

agaaggaggt ggaccgaagt cttgaccatc tgccttcctt ctcatctgag gcgcgaagtt 27120

attgatcatc aaatgcagct aatgtcactg aaagaatgac aacacaacat gggtggcttt 27180

gctggaagta gccacgtgtc ctgggaagcg agtttgccaa tgtccagcct gaaccccctt 27240

gagcctcttg atgctagaac catttttcag gaaaagagag gttgagggaa gcactgggtt 27300

cctgctcagg gaggtggcgc ctgcatcccg actgtgggag gtcccgggac cagctctctg 27360

ctcctgcagc tcctgccttg ccagggaaag ctagagcaat ttgggagaca cctgcagatg 27420

gaatgggtct aaaaacacat tcaccaatga taacgtcaag tattaatcca catagaaatg 27480

aaattaaatc acaaatatgc aacagcacca caaagtcgac aaaaatagac acaatgatat 27540

tcacaagaca tttgataaga ctctcattaa ctatttttga tgtcaagaaa tccttgatat 27600

tctctccagt tgtgtgtgtg atgactagca tttgcttcac acccttgctt gaaggagtag 27660

tttagttatt ctgctgtggg tgggtccgca ttatctgaaa atggatgatg ctgcggactg 27720

gctgatggga accttggagt cattactatt cttctgtctc cctatttttg tgttcagaga 27780

tcttcatcat aaaaatgtac aacttataga gagggtgaag cttgggacag tgtctcccat 27840

gggcttgaat tttcatagct gtgaaatggg tcaaacagag gctgagggac tctgggcacc 27900

tgaaattcaa ttcagctcac ggctgtgatt tggggcaggg gtggctggga ctgagctcca 27960

aaggggcccc tctctttgtg tatcattgtg gaaaaagccc ccaggccttg cagctctgtt 28020

cctgtcctgg ctggccatcc agcagctgct gccatgtccc tgatgaacca gaaacagtgg 28080

gggtcccagc ccgggcacta tggattaacc gagcctgtca aggccttcaa gggaccaaat 28140

tgcccactgt cagggacccc gggatgaggg tgtctctgag tgaggctgcc ggtgcaccat 28200

ggtgcagggg gtgaggggac ggaacaacgt gagtgacaga gggagtgggg cctggtggga 28260

tgggggtgtt tggggcagtt gagatcccgt gaaagcctga ctttgcctat agatagcccc 28320

taggtcatgt tcaccctgtg ctgaccttgc gtgacccagc cttgactcta gtccagaatg 28380

agactctgac acagactctc cccgacaacc atgggatgcc cgttctgtgc ctgggatggt 28440

gtctttgctc atttgtctca gtccacagga gtcctcccct cacacgagct gaccatcaca 28500

ggctcagcca cacaccaagc acatactcac tccaccccag agcctgactt tagaattggg 28560

aacttcatgg ttgagatctc agccacgtgc tggttaactt tgaacctcac cctaagccca 28620

tctgatcata actgatcacc cgtgtgcccc agcctcaccc cactttccag agctagcgac 28680

gtgaacacca gtatcttgca gagcatccta gggctcacta agttggctgg gatggtgagc 28740

aggattctgg acttcctgtt tccctattgc aggacttgta agtgtcttta cttagccacc 28800

ctgtattgtg aaaatctgtg aaattcacct gtgagtttcc attcccagac ctttccacaa 28860

gttcctggtg atggcaccac catggacttc tcacccgata aactgggcac ctggattttc 28920

ctcatccttg cttagaattt aaggaggttg caaactaaat atgagataag aaatgaagac 28980

agacagcagg atttctggtt gacaatgagg atatttattg agggttactg ggtgcaggga 29040

gaagggctgg atgacttgga gtgggagggg agacacatgc cctgatcctg cagctctagg 29100

cccccgtggg taggtgaggg tcggggacct aagaacattc tgcaggggcc actgtcttct 29160

ccacagtgtt cctttcatgc gtgacctggc agctgtagag gttgtgggac ttccactgtc 29220

gggtgccagt cttaggtagc tgctggccgc atacttgttg ctctgtttgg agggtgtggt 29280

ggtctccaca ccctgggtga tggtgatgcc atctatcttc caggccactg tcatggctct 29340

cgggtagaag tcactcatga gacacaccag tgtggccttg tcttggagct ccctgcctct 29400

ggggtcagag gagatgaagg tgtctcagtg accagctgag gcatacaggg gtctgaatag 29460

ggaggagaga gatcagcctc ccaggaccga ggtcccaaag gaaaccctga ccacaggctg 29520

tggcaagtca cagacaaaga tcgaaatcag aataaaattg tcaccaactt ggcagggaaa 29580

gaaagggaag aagagactca tctaaaatga tcagctgggt tcctccacca aatacaaaat 29640

gcggtgacac aggttcatac aaaaactctt ccctggggcc cctaactccc cctccccatc 29700

ccccacctga agctatggtg caggaagcca gaccactgcc cctggaaggc atttgctcag 29760

cagagtccac acacctgcct gcccttctca agctgagatg ggacagtcga ccaagcgtat 29820

gcagtaacgg gacgtctgga aatggcgagg cctgaggagg ttggtgtctg gatcaggttt 29880

tccaggtgac tgtgacccag gaatgggaat gagctccttg agggcatttg cttcacgaag 29940

gagtgaagtg acatggaagg ggacagggat tctctatagg gttgttgtaa ccctgatctc 30000

ttggtgtgac tacagggtaa gagcaggagg cgtcctcagg tgtccccagt gagtgcctta 30060

gaggaggggc agcttgcccc agagtcagca gtcatgggaa gctccaacgt gaccacaacc 30120

acaactccag ggcgtctcac ttccgtcctc cttgtgttgt cttagtttgg gatgggtctc 30180

actaggtttt agtatgaaat gccagcctca acctccaggg agaagttgga agcaagcccc 30240

tgttaccctt tcagggtctt gtctcatttc tgggtatgtc tggagtgtct gactcaaaag 30300

ggctgtgtga gaagcacccc aagtgtggtc actaaatccc acctcccagt tatggggact 30360

agtccacttg gggttaatgg gcaattccag gcacaaggca ggagaaagat gtgcccagga 30420

tagattgtga acagagagcg gaaagccatc gaggtctcct gggttccttc aggaggatcc 30480

tggggccacc agaagaggca ctgagtgggc cacggtgggt cattgtcatc atcaatgaaa 30540

tagaaccatg gtggactcaa gtgagtcaca tacagtaaca taactttgag ctcaccatga 30600

aataattgtt catcttcacg agatggcagg caacatggtc atcattatta agatgctaaa 30660

tgaaaggaaa gactcaacat gtctatgcta tttcttactg aaataagaat cacactccca 30720

gaggagcaaa gaacatatga gggcacacct tttacagaca gtcagtagaa ggaaaaggtg 30780

gtcgaactga aatgcagtca ccttgcatcc atataccatc taatggcact ggatacaaag 30840

tgtgaagccc agctaacgtc acaaatagat gaccccacat gggagcccag tagccaagcg 30900

gtcttgctga aaagtcacac tgagccttgt taaacctttc tggccaagaa ccaattttca 30960

ggaaacgcag aagcagagag gagcgcttgt ttccagcttg gggaggccac aggctgtatc 31020

ctgaccatgg gaaggtcctg ggaccaaccc ccagctcctg tagctcctgc cttgcaagga 31080

aattaaaaat caatgaggaa gatacctgca gacagaacag gtctaaaaac acagtcacca 31140

gtcacaattt caaaccatat tgaatcctca tagaaatgaa atggaaaacc acagaataac 31200

aacaacacaa acacaggtct agtgaaagaa gcataggata tttgtgaagt atttgatggg 31260

aaacattctt tttttgatat gaaagaatta gatatttata tttaggtttg taagaatgta 31320

gttttttggg gggggtccat atttttagag acacaaatgg aatatttaga ctaatggaca 31380

tgatgtctac aatttgcttc aaacaactgt gtaaaagagg ttacttgatt gtgggtgggt 31440

caacatgagc tgtattatcc tgttttttta ttttcttgct gtcagtttcc gtaatatctt 31500

acaaggcagt aattagagag attagtcttg ggacagcacc aggggacatc cagaggggat 31560

cctacccagc agggccattg agaaggtgga gatcaaagcg gtcctctttg ctataggggt 31620

ctcagcgttc acatctatga aatggggcca gacagggtgg tgggaatcag gacccttaag 31680

tttccttcta acttgtaggt tgtgacttga ggcaggggtt ctatgattag gctccaagag 31740

gaaccccttc tctgccccac tatggcccta gagttctctg cccttgcagg ctctgtctca 31800

ggcctggcct ggccacccag cagctctggt gaaggcacct gccacaccgc tgatgtgctg 31860

gaggcaccaa gaaggggcct gacagtgccc aagactgatg aggagggctg ggaagtgcat 31920

ctcccaaggc cacctcactc gctgtcaggg accctgggat gattgtgtct ctgacatcag 31980

ggtgtaggtg cacatggagg ggctgaacca catgattgag ggagggagta ggccctggtg 32040

gggaggggtc taaggtagct gagtcccatg aaaacttgac ctgcctgtag cccttaagtc 32100

acacttaccc tgccctgacc tagtgtgacc tggccctgat tcttgggcga gtatgatttt 32160

gacacagcct ccatcagaca accaatggat gccgattcta tgcttgggat ggtgcctttc 32220

tcatttgacc 32230

<210> 73

<211> 115

<212> PRT

<213> Intelligent

<400> 73

Met Ala Trp Ile Pro Leu Phe Leu Gly Val Leu Ala Tyr Cys Thr Gly

1 5 10 15

Ser Val Ala Ser Tyr Glu Leu Thr Gln Pro Pro Ser Val Ser Val Ser

20 25 30

Pro Gly Gln Thr Ala Ser Ile Thr Cys Ser Gly Asp Lys Leu Gly Asp

35 40 45

Lys Tyr Ala Cys Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Val Leu

50 55 60

Val Ile Tyr Gln Asp Ser Lys Arg Pro Ser Gly Ile Pro Glu Arg Phe

65 70 75 80

Ser Gly Ser Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Gly Thr

85 90 95

Gln Ala Met Asp Glu Ala Asp Tyr Tyr Cys Gln Ala Trp Asp Ser Ser

100 105 110

Thr Ala His

115

<210> 74

<211> 32230

<212> DNA

<213> Intelligent people

<400> 74

gaattcctga cctcgtgatc cacccacttc agcctcccaa agtgctggga ttacaggcag 60

ccactatgcc cggctgtgaa catgaatttt taaactgcat ggtgctttgg gtcccagctt 120

ctccatctgt acaatgggga cagtactaag gttttctttt tcttctcagt tgactgaatt 180

atttccatag tctgtctgtc ttgccactct tgttacccac acgaaaggac ctaaggtaat 240

ttctgacagc ctgggactcc ttgtgcaaaa cagaaggtgc cacagacctc attttaggag 300

aaacctttgt tttcctcttg gaaccccaag aactgtaagc agacaggtcc atctcaaaat 360

ccaaggctct gctctgtttt gctttgcttt acctgacttt ttttttgatt tgggtgggca 420

tcaaaaatta gtaggggaga gctaaagaaa gttgtggata tgaacatgta tttatggtaa 480

ggtaagttat gaaggagaga aatcttagga tcttgtatgg caaattcttg cactaaagta 540

gataatgact aattaggaaa gcaggaaata taggacaagt cagaaagtca taagatggga 600

gagaaaagct tacgcctggt agatcctctc ctgtctagaa gtgtcgtatg tgtgatgttt 660

atatgaagga gctctaatta attggattaa aagataatga aagctctaaa attgtcagaa 720

aaatagaagc tctaatgcct tttatttcac gtgacttcag caatctttgg gaaatgagag 780

ttttaaaatt attggtggta aaaagtcttc aaaaagtaga catttggtct aaattaagtc 840

agaggttaaa ttggctaagt gctttaatgt cataaactgc ttctttgact ttggaaaatt 900

gttcagttta cctaatttgg aactgttaga tttctaggta aggcctgggg agaggtggag 960

ttagccatgt ctcctagcta tgctggaaga gttagacttt atctgcagtt ctgtcatgta 1020

tcctggactc tgcacctggt atgtaattga aactgcttac attgaaaaga aaaattatgt 1080

gtttttggtt ttaaaagggg tgggaatatg gttttttcca aaaagtgagt tttggcaaat 1140

ttagaaagtt taagaattat ttttagttgt aagaaaataa agtttaagca agttatagaa 1200

ggtttatgaa aattaacttt gtaaagaatt gtgtgtgtga gcaagttgac taaagggaca 1260

gtaatgttag atgtaataaa tgcaggtacc cttccacttt atcttctgtt tcttaggtta 1320

taaatattca ctacctacgt ttctaaaaac tctttcactg taatattaac acataactcc 1380

attctccttg tattgcctgc tttttcatag tatcctgaaa ttgcattagt agatttagat 1440

tcagaagatg gtggggaagc atcttatgag tgttttaagt aaatttaagg caaagggttt 1500

tctgtacatg ggtggttatc acagcatcac ttgttgcagc aaaacactgg gtaacaacaa 1560

tgtttatcaa tgagaaattg ttcggataaa tgatgatttg tgccttgcga ggctcaacta 1620

ttcaaaagca ccctcttttc ctcacttgct ttacctttgg ttctgggcct ggagcctttc 1680

agaggttact caggacagtg gactcctgtt tcctgtgact aatctggaca gcggcttcct 1740

atttcattct tcagcatctt ccaaaaattt ctgaagataa tttcttcctt tgtgaccacc 1800

catactcttg gtttactttg cagttaacaa tttcatgcct tcactggatt atctgtgtga 1860

aggtgtgggg tgtatgtgtt catactctgg agggagcaga ggaaaatagc cacagccaaa 1920

ggagtaatct tgcactaaat actcattata agaatgttga tacgttcata gatggtaaat 1980

ctgtcagtaa tttttacttg tatgattctt ggttaagact gcactcactg gtgataagcg 2040

tgctgaaggg atcataggtg ctatacttat gtgtgggaat tgcattcaaa ccaccaatgc 2100

ctgtctactc aagttttgtt gccttttgat taaaaactct tgggaatgtt tccctggcat 2160

gcaaatatgg gaactgcagt ttgattctag gtagaaactc tttggtagtt gaaatgccat 2220

aataagagac acagctgtca atgggcaaca attcactgtt ggtaaggaat ttacagtgct 2280

acttgccata ttcttcttta tccagacttt gtgccaagaa aatgaaattc tttttaaaaa 2340

tttttttcta gagtaatcct acaatattgc tgttatcaag cctgatgctg taattagtag 2400

aaaatgtcta gaaattaaaa gaaaagtagt attttccaac tatggttaaa ttaattgcaa 2460

aataacttcc tcttaattgt cagattggag acaagcttat taagagtgaa ataaaacata 2520

atgtgttgct ggtgctaggt acctctaaag cctttgagca ctttcttctt tataaaggct 2580

caatctactc atgtgtcctg taaataatcc tgctctacca gcatcctatg taaattttaa 2640

aaattatagt tcatatatta gattgtaaca aatgttgttc ttctttaatt atatgcaatg 2700

caatcttgtt atagactcac attagcttgc cttattccct gtatgtttgc tgtaaaatgt 2760

atagtccttt tcaatatatg atagcttaaa gatggactta atattttgtt ttagaaccaa 2820

aatcttttat cttatataca ggaaaattac attggaattc agagatcagc aaacatttcc 2880

tgtaaaggat gaaatagtaa ctattttcag gtttgctcag catgtggtct ctgacaacta 2940

aactctgaag tcataccaag gcagcatcag taatgtttaa atgagtgaat ggcgctgtgt 3000

tccaataaac tttatttaca gaaacaagtg gtgggccaca ttttgactgc agggtatagt 3060

ttgtggaccc ctgaacaatg gctttttctg aaacaacaca attgaattga gatttgaaag 3120

aatgttgagg cagcattgtt tgtctggggt aatacctgag gtttgtcatc tcacaccagg 3180

gaaatcaggg atgtggacac acaagaagtg agttcaagag tggatgatta ataggtgaaa 3240

gaaagagaaa agagaacagc tctctctcct gcagaaagag agaggggcac ctgagtgggt 3300

cttccggttt tgtggtgaaa agcacaaggt tttataaagg agcttgagga ggtggtgtct 3360

tgatttacac agggctctag agactggttg gaccaggtgt gaacgtttgc atagcacagg 3420

gagaagctgg ccattgcacc ctaatctttt attatgcaga tgggttcttt accttaccag 3480

gaccatattg tctgttcctt actgtacctg tggttgacag agaaaaggga agatggagcc 3540

accatgttga acatgcctgg cccacaggta gccttttcct attggcacag ctgctggcat 3600

tcctatgaga gcttatcaat gtctgcagcc caattttaca agatgctctt tgttagaaaa 3660

taaatgatat gggttttttt tttcattagt aggaaaactt tacactagga cttccttact 3720

ttcagtatct gcctaaataa tttcttttta attcctacat caatgttgca tgtgcaatag 3780

agaaggtgac atgtagggag ctgtgtcagc aggggttgca tatttcttga tgagcatccc 3840

atgatgcatt cttcttcact tgtggcaaca ttagcattaa ctaatctaca gtaaagtaga 3900

tgggtccaca ggtcatggga catacactag ctctgccaat tacatgttga tttaaataat 3960

atcattctct aaggacagct ttttttcccc cagatcctgg tatgtatgtg gagttgggga 4020

gagacaccaa actgtacccc atttcctcat acattgctta tacttttaga agtgtttgaa 4080

agataatatc acatactagt agtttactgt ttcttattca agttagtttt aatagtcgct 4140

agggtgaatg ttttcaaaat tgtatagata ttcgggttga ggcaatacta taaattatga 4200

gttcatctca gtttttttgt gctagcgtaa tgacatttcg gagacataat gcatatgtgc 4260

aaaatacttc acctctctgg aaaacagtaa agggctacat aataggacag acaaaaataa 4320

aggcttggcg taatggctca cacctatgat cccagcactt tgggaggaca aggcgggtgg 4380

attgcttgag ctcaacagtt caagaccagc atgggcaaca tggcgaaacc catgtttaca 4440

aaaacgataa aaataagccg ggcgtggtgg catgtgtttg cagtctgaga tacttggggg 4500

actgaggcag gagaatcgct tgagcctggg aggtcaaggg tatagtgagc cgagatcgtg 4560

ccactgcact ccatccagcc tggataacag agtgagaccc tgtctcaaaa taagtaaaaa 4620

tttataaaac aaagaatgtt tctaactgta ataaaggaga agtgatcaag cttaagtgta 4680

gcttagattt aatctttgat ttccgggtac tatattactt aaaaatatga tttggatatt 4740

tagaagttgt tcaaattaga aactcaggag tggggtcact ggtcactaag gtgctgaata 4800

tttttaatac cttagtggat atgtaatcaa caaagccaac ggaaccaata gagcatttgc 4860

tttcataaga atttaagtgt tgctgggtac tcttaaatcg agctcacatt attggtgtca 4920

tttaatataa ttactgaatt tctttgaacc cagtttccac tttttaaaaa ttatttagct 4980

cattaggctg aaaacttctc tttgttacat tttattacac ttcagtgtaa tatgtgtaag 5040

atgtacattg cttctctcta gaaaggtgag acaagttgaa gcagggttgg gggtgggtgc 5100

ttccagttca taggtaaata ggagacaaac ggttgcattc ttttgagttt ctgattatcc 5160

tttcactgaa tacacaattt acatgcaaga tgccatagag gaatagtcac ttatgcctga 5220

gtctggctta gtgaatctgc atttttacat aaaaaactgg gcagagaaag ccaatcagat 5280

acacatttgt ctcaggtgag gagagggatg actttgggtt ctgtcctttg tcccccacct 5340

gtaaagataa tctatcaatt gccaatttat attgccaggg tcaaattcaa cagaactgtt 5400

ttagtgtaaa gatcttgagg cccacaagaa atttcctttc tgacaaatta tgagggaggt 5460

atatagcttt cgaaatcttt gtagttattt ttttatttta ttatttgggt tttttttttc 5520

tttattgaca aataattgta tatactcaca gggacatagt gatgtttcaa tacagaaaag 5580

ttatagtgag cagagcagga taatttcata tcctttgtta aaggaaaata aatctccggg 5640

acaccccact aagccaaata atagttatct tatttagaaa taaaatggga ggcaggtttg 5700

tttgatggag ttcctagctg aggtgggaga attgcttgaa cctgggaggc cgaggttgca 5760

gtgagccgag atcgcgccat tgcactccag cctgggcaac aagagcgaaa ctctgtctca 5820

aacaaacaaa gactatagaa aagctatttg tgaaaatgct ttttgatgtg ctgttttatg 5880

tcacagaatg caacctgtgt tttgattcag caagttcaaa acactctttt tgtggaatct 5940

aagaattgat tattttgatc ctattgagct ttcataagaa aatatgaata tccagttgta 6000

aaaactagaa ataagctata agctatctgt ggaatgcttt gtgaggtgct gttttatatc 6060

ataaaattga acttgtgttt tgattgaaca agttgaaatt actcattttg tagaatctaa 6120

gaagtaacgt tttggaactt cttgagccct tattaaaaaa tagaaatatt cagtcctaaa 6180

aactagaaac aagccatccg tgcaaatgca ttgtgatgtt ctgttttgta tcataaagtt 6240

gaacctgtgt gttgattaaa caggttctga acactttttt ttgtagaatt tatgaagtgg 6300

catttctgag cctattgagc ccttatacaa acatacagtt ttaaaagcta gaaacaagct 6360

atctgtcaaa atgctttggg atgtgctatt ttatgtcacg gaatgaaacc tgtgttttaa 6420

ttcaataggt tcaacacatt cttttggtag aatctgagaa gtgacattcc caaacctctg 6480

aggcctttaa aagaaaatac gaatattttc ctctaaaaac tagaaacaag caatgcgtgg 6540

aaacgctttc tgatctgtta gtgtcacaga ttggaaactt tgttttgatt gagcaagttc 6600

aacgcactct ttgtaggatc taagtagtga catttccaaa cctattgagc ccttataaga 6660

aaatatgaat atttagtcct aacatgctat ctgtgaaaat gctttttttt tttttttttg 6720

gcacagagtc ttgctgtgtt gccaagctgg agtgcagtgg tgtgatctcg gctcactgca 6780

acttctgcct cctgggttca agcaagtctc ctacgttagc ctccctagca gctgggacta 6840

caggcgtgca ccaccacgcc cagctaattt ttgtattttt agtagagatg tggtttcacc 6900

atgttggcca ggatggtctt gatctgttga cctcgtgatc cgcctgcctc agcctcccaa 6960

agtgctggga ttacagacgt gagccaccat gcctgactga aaatactttt tgctatgctg 7020

tttcataaca cagaattaaa cctgtgtttt tgtgagacag gttccaaaca ctctttttgt 7080

agaatctaaa aaagtgatgt atccaaacct tttgagccat aataagaaaa catgaatatt 7140

tagccctaac tgctggaaac aagctatctg tcaacacact ttgtgatgtg atgctttata 7200

tcacagaaat gaacttatgt tttgatgaaa cagattcaaa acactattgt agaatcttaa 7260

gaagtgacat ttctgagcct attgtgacct tataggaaca tacaaacatc cagtcctaaa 7320

gtctacaaac aagctatctt ggaaaacact ttttgacgtg cagttttata acacagaatt 7380

gaacctgcat tttgataaaa acacgttccc aactctcttt ttgtagaatc taagaagtga 7440

catttcgaat ctattgagca tttacagtaa cgtatgaata tcttgtaata aaaactagaa 7500

acaagctatc tgtaaaaaca ctttgtgatg tgctatttta tgtcacagaa tggaacctgt 7560

gttttgtttc agcaagttct gaactctttg tagaatctaa gaagtgacgt ttttgacctt 7620

attgagccct tataagaaaa tacaaatatc cagctataaa aactagaaac aagctatcag 7680

cgaaaacact ttttgctgtt tcatatcaca aaattgaacc tgtatactga ttaaacagtt 7740

tccaaacact ttttttgtag actcttaaga agtgatgttt ctaaggcagt gaggccttat 7800

gggaacatat acataatcca gtcctaaaaa ctagaaacaa gctatctgtg ataaccattt 7860

gtgatatgct gttttatgtc agagaatggt acctgtgttt tgattcaaca actgtgagac 7920

actctttttg tataatctaa gaagtgatgt caccaaacct attgagtttt ataaaaaata 7980

tgaataaccc taaaaacgag aaacaagcta tctgtcgaaa atgctttgtg atgtattttt 8040

ttatatcaaa taatggaacc agagttttga tgaaacagct tccaaacact ctttctgtag 8100

aatctaagaa gtgacatttc caagtcaatt gagttcttac aaaaaaaatg caaaatccag 8160

tcccaaaaaa ctagaaacaa gctacctgta aaaatgcttt gggacatgca gttttatgtc 8220

acagaacggt tcctgtgttg attcaacaag ttcaaaacac tcttgatgta gaatctaaga 8280

agtgctgttt ccaaacctat tgaactttta ttaaaaaagt aaaaatatcc atccccaaaa 8340

actagaaaca agttatctct gaagatgctt tgtgacgtga tgttttatgt cacaaaatgg 8400

aaactacgtt ttgatttagc aagttctaaa cactttttct agaatataaa aagtgacatt 8460

tctgagctta ttgagctctt agacgaacat acaagtatcc actcctaaaa atgagaagaa 8520

aaacctatct ctgaaaatgc aatgttatgt gctattttac atcacagaat gaattttcaa 8580

acactctttc tgtagaatct aagaagtgaa atttccaaac ctgttaagcc cttataggaa 8640

catatggata ttcagtctta aaaactagaa acaagctata tgtgaaaatg cattgtgaag 8700

tgctgtttta tgtcacagac tggtccctgt gttttgattc accaaagttg aaacactctt 8760

ttggtagaat gtaagaagtg atgtttccaa acctattgag cttctataag aaaatatgaa 8820

tatccagccc taaaaactag aaataagata tctgtgaaaa cactttgtga tgtgctgttt 8880

tatgtcacag aatggaacct gtgttttgag ttgacaagtt ccaaacactc tttttgtaga 8940

atctaacaaa gtgacatttc caatcttact gagcccttat aggaagatgt gaatatccag 9000

tcctaaaaat gagaaaaaaa gctatgaaat tgcattgtga catgttgctt tatgtcacaa 9060

aatggaatct ttgtttttat tcaacacttt caaaactttt tgtagaatgt aagaagtgac 9120

attttcaaac ctattgagcc cttataagaa aatgcaaata tttagtcata aatattagaa 9180

gctctctgtg aaaatgcttt gtgatgtgct gttttatatc acagaattga acctgcattt 9240

tgatgaaaca gcttccactc acttttttat acaatctaaa aggtgacatt tcctagcctc 9300

ttgagccctt attggagcat atggatatcc tgtcttaaga actagaaaca agctagctga 9360

gaacatgctt tatgacatgc tgttttatgt aacagaatgg aacctgtgtt tttagtcaac 9420

aagttccaga cactcttttt gtagaatctt aggaagtgac attttctatt tttattgagc 9480

ccttataata aaatattaat atttagtcct aaatattagg aaaaaaccta cctgtgaaaa 9540

cgctttgtga tgtggtttca tatcacagaa ttgaacctgt gttttgacaa agcaactttc 9600

aaacactgta gaatctaaga agtgacatta ctgagcctat tgagctttta taggaacata 9660

caaatatcca gttccaaaaa ctagaaacaa gctcttggtg aaaacgcttt gtgacatgct 9720

gttttatgtc ataaaaatgg aacctgcctt ttgatgaaac agcttccaaa caggtttttt 9780

tttagaatct aatagttgac atttctggcc ctattgagcc cttatagaaa catatgaata 9840

tccagttcta aaaactagaa gcaagccatc tgtgaaaatg ctttgtgatg tgctgctttt 9900

tgtcacggaa taatacatat gttttgattc aacaagttca agacactctg tagaatctaa 9960

aaagtgaagt ttttgaacct attgagactt gattaaaaaa tacgaatatt cagccctaaa 10020

agctagaagc aagcaatctg tgaaaacact ttgtggtatg ctgttgtatg ttatggaatg 10080

gaacctgcat tttgatgaaa cagcttccaa aacacttttt tgtagaatct aagaagtgac 10140

attttggagc ctattgaacc cttacaggaa aatacgaata tccaatccta aaaactacaa 10200

acaagctatc tggtaaaacg ctttctgata tgctgtttta tgtctcagaa tggtactggg 10260

gtttcaattc agcaagtttg aaactttctt tttatggaat ctaaaaagtg acatttgaga 10320

acctattgag ccgttataaa aaattcaaat attagccata aaaattagaa acaagctagc 10380

tgtgaaaacg ctttgtggtg tgctgtttta tgtcaaagaa taatacctgt gttttgattt 10440

ggcaagttca aagcactttt ttttctagaa cctacgtagt gacatttaca aatctgttga 10500

tcttttataa gaaaatatga atatccagcc acaaaaacta ggaaaaatct gtgaaaatac 10560

tttgttattt gcattttatg tcacagaatg aaacctgtgt tttgatgaaa caacttccaa 10620

accctctttt tgtaaaatct aacagtgaca cttatgagcc tatggagccc ttatagaagc 10680

actcaaatac ccagttttaa aactggaaac tgtctcaaaa acactttgtg aggtgctttt 10740

ttatgtcaca gaatgaaacg tgtgttttga gttgaacaac acacaagttt ccaacactct 10800

tgaatctaag aagtgacatt ttcaaacttg agcccttata gaaacataca aatatccagt 10860

cctaaaaaca agcaaaggct atctgtgaaa atgctttgtg atatgctgct ttgtgtcatg 10920

gaatggaatc tgtgttttga ttcaagtttg aaacactctg tagaatctaa caagtgatgt 10980

tttgaaccta ttgagtcatt ataagaaaat gcaaatattt agccataaaa atgagaaaca 11040

agctttttgt gaaaatcctt tgtgatgtgt tttctatcac agaattgaac ctttgttttg 11100

ttgaaatagt ttctaaacat acttattgca gaatctatga agttatattt ctgagcattt 11160

gagcccttat aggtacacat gaatatcaga ttctaaaaac tagaaaggag ctatctttga 11220

aaactatttg tgatgtgctg ttttatgtca caaaatgata accatgtttt gattcaaaaa 11280

gtttgaaaca ctctgtagaa tctaagaagt gacatttctg agcctattga gcccttagta 11340

aaaaaacaaa caaaaaacga atatccactc ctaaaaacca gaaaaaagct atctggtaaa 11400

atgctttgtg ttgtgctgtt tcatgtgaca gaatggtgcc tgtgttttga ttaaacaagt 11460

ttgaaacact ttttttttgt agaatataag tagtgatgtt ttagaatgaa ttgagccctt 11520

ataagaaaat gcaaatattt aaccataaaa ataagaaaca aacaacctgt gaaatcactt 11580

tgtgatatcc tttttttatg tcacagaatg gtacctgtgt tttgatttaa caaatttgaa 11640

acacactttt tgtagaatct gagatgtgac atttacaaac caatcttaag ccagttagat 11700

actctgatga gccagctgtg tgtgtatgtg tgtatgtgtg tgtgatcttt gattattaaa 11760

actgaaaaaa taatcgtata ttcatgacta gtagacacag tgttttaaaa ctgattctga 11820

aagatttgag ggttaggctt gttttaaaaa caattgcata ggtcctggaa acaccaagaa 11880

ttagtggctt ctcattccta gtcacatgaa aaatatttta ctactcttgt gaatggccat 11940

aaataatatg cagaattttt tctgtgtcct aaccttttat ataattggtt ctacagtgta 12000

catcactgtg tgcaactttg ttttgtgtgc tcagacttac attttccagt tttctccact 12060

ttgatacaat tggtctgtgt ttcatacatt ttgtattttc tatagcgttc ctttatatga 12120

ctctaccaca ttttattgag tctcctcatg ctgacaggta agtagatgaa ttaataaatg 12180

aatgaataaa taaaaatgag agcaaatttc aaacaggtga ttaatttagt gattgtaacc 12240

atggactcag caggttagaa ataaaaactg aaagatttag aatgcaagca ggcacatcat 12300

accttctaat agccattgga actatgaccc acaaaccatt agcctagaat gatgggacat 12360

atcttgtctc taaaaaaagg tcactgttca gttctcaaag aatggatata aaaatggcag 12420

ataccattcc acattattct acttccttta acctctacgc ataaaacttg agctgcaggg 12480

aatgttacct cctgttctta aaaagacttg tagttcagtg aggaactcac tcctttcttc 12540

attcatgtgt tcaaataata tagggaaatg aatggaaaag tgaagagaag ttgaagacta 12600

ttatggggat agcacaggat tagtcagcct catggtatta ttgatctgcc ttattccaca 12660

aatatttacc aagcacctgc tacatgcaat ttgtatacta ggtgctgggg atacaggaga 12720

taacaaagtc cctgctgtca atgaaagggg tcagtccttt gattcatctt tagttaattt 12780

taagcacaca acaactatct caaactttct tcacactttc caagctcccg atccttcatg 12840

cactcttggt agatgcctga ccttacctct taccttacta gaacctaaga ccacctggta 12900

taaaatccct taagcacact cagatttgcg tgattttaca atgatgtctg ttcccaaagc 12960

atcctcactt ctcttggagc tgagggtgaa gagctcctct ttcttcccat ccgtgatctt 13020

ggaacatgta cgactacact tgtgacagta aatgcttaat acaggcagaa ggtttgttcc 13080

ttatttcctc cctggggcct ccctccagtg aatatctctt ctagcattgt cagttgtatc 13140

caacagaaaa gcagagatct ctccattggt ccaaaaacct aagatggact ctcactgcta 13200

aacttctcaa atgaaaattc ctccacttta gtcctgatta ccactcaaac actgttagac 13260

tgggcaggtc actacccctg cctagttctg tttccccatc tacataacag agaaataatt 13320

gcgccttatc tatcctacag aagtgtgaag aggtttgatg tgacagcagt gaaagcaatt 13380

taaaaagcac agagtgctat acataagaag acattttcat aattattccg ctctaacatg 13440

atatctgtat tcatacgatc tcaaagtact tacccaagca tggattctaa ctcttgtgta 13500

tgtaagcaaa ggtgtaagtc tgtcaccctc catgcaaata gaatctcccc tcccttggcc 13560

cctgcatctc accatataca ggaattaact caagatgaat caaagtttta aatgcaagac 13620

cgcaaactat gaaaatcctt taagataacc taggccatac ccttcttgac aaaggctttg 13680

gcaaagcatg tgtatgtcta agtccctaaa agcaatagaa ataaaaacaa ttattgacaa 13740

gtgagaccta atacatgaaa gagtggctgc acagtagaag atattaccaa cagagtaaac 13800

agacagcctt cacaatggaa gcaaatgttc ccaaactaca tctgacaaag gtctaatatg 13860

tacgatctac cttaagtcct taaatcagat caaaaagtcc aattaatggt tagagacatg 13920

aatacacact gctgaaaaga tgtaacagcc accagtaaac atgaaaacat gctcaacctc 13980

gttaatagag aaatgcaaat caaaagcaca ctggtcaaaa tggtgattat aacacagtcc 14040

acaagaacag ataccagcag gacagaggag ggcactgggg caattgatca tgattcttat 14100

ggatagtcat ttagtgttat taatgaattt cataacaaaa tgatcaccat acaaaaaata 14160

caaaatgtgt tttcttccct gagaaaggaa aataaaaact tggaatgccg attcactatg 14220

ccaaaaggaa aagttaagct gaaagctgag tcatgcaaga aactgccttt cttttttttt 14280

cctgagcaga cagctactga taaatagtta aatttctcca caggtagcta ctctatgttc 14340

atctcatctt acttaaagtg ctgatttact gagcaggaga aaaatatata actgactatt 14400

cctttacctg tttgttttct cttgcaacat gtggattcag caacgtgacc atactctccc 14460

tcttttcccc gcagcctgct tttctcttct aaacactgaa accctaaaat atttttggag 14520

aaaggcacag acctctgtcg cccagccatg tctttaacct tggaagaaga aacttctaaa 14580

ttgataagag atctgtctca gacacttttt cgtttacatc ccgttatctc agaaagcaaa 14640

acaggctttg ttccaaccca aagtaatttg tgaatattca agcggtatca ttctgatgta 14700

aaatttacat aaattcttgg gtttaccaaa atgcaaatgg gcaagcagat cactatttac 14760

agataatggc ctaaaaaatg gtaagagtaa atgctgtgga gctaaatgtt tctgagtctg 14820

ccatttccta gttttgtagt ttctctcggg caagttaccc aaagttgctg tatgttatta 14880

tcttgacatt taaaatagaa gtaataactt tattataaat aatagttaat accatagggt 14940

cattaggaaa gtgtctggca tatatagtca caacccagtg aatgccatta ttgtttttgc 15000

tattatcagt ataataattg atactaattt tgatgctgag gtaaaataat atctggcagc 15060

ttttaatttg ctaatgttaa ataactgaaa atagctttaa aaagtgaaaa tatccaaatc 15120

aatgacaata tttgatttat ataaaataag cttcattttt tttagttttt ttgctcctaa 15180

ttattacttc actcatcaca ctgatatctg agcacttttt aaaacatatc cacccttctc 15240

agttgtacta cttattccct tttcttgtaa tcccagcctt caaagatcct ttctcatttt 15300

tctaactcat ttcacgactc cctcaccttg acccccagtt atatctctct tgaagtcaat 15360

tccattgaat tgactacttg gcccaaactt caccccaact cagtctctct tcaggaagga 15420

aagtccccac acaaaaagga atgtgtcttt caactcatgc gccctttttg aagaagcttt 15480

ccagacagct tccttacagt gatctccgtt atccatcctt tagatcgctg tggagtccta 15540

caggggcgca atgtggacag tcattcaaat gaagcacacc taagtgcttg cattttctaa 15600

aggcccattg attcatgtaa ctcccgtact tgcacagata agatgttcct gcctgaatgc 15660

agcagatgga aatgaaaatt gtattctctg tcaaggctga ttgtattaca gcttaatgat 15720

ttgcatttta aaacttggaa gttctcataa atattaaagt gaaagaacaa ttggctacat 15780

tattaggtag gttcggctgt tcgtataacc tggtgctaac tatctaacct gcctgagtga 15840

gcctcagtct tcattactgc agcatttgta agatatcaat gcctgcttga actcatgtct 15900

tactaagatt atggccagga ttgagtaaca tatggatata aaagcacttg acaaatcaga 15960

gtatttcata aatgtaataa taatatgcta catttataac atctatactc ttttcctgtg 16020

tctatcttca tttaaacctt gcagtttcta tcataatcaa agattttgta atcaggttac 16080

aatgagtagg atttaaccca taagcaattt tcaaatttcc tagatatgtt ttcagttttc 16140

acatttatgt tggtgacaaa attctgtttt cagttttggt gtttgcatct tctaaaacag 16200

tgtcttgggt acttgctctt tgatgagaca ctgtgcagac ataaagagtg taagaaaaca 16260

gaacagttta tctcagcttt ttgtggtcta gataacctgc acactgcttt cttggctcac 16320

tgcaacctcc tcttcctggg ttcaagtgat tctcctgcct tggcctcccg agtagctgag 16380

actacaggca tgtgccacca cgcccagcta atttttgtat ttttagtaga gatggggttt 16440

caccatgttg gcccggatgg tctcgatctc ttgaccttgt gatccccctg ccttggcctc 16500

ccaaagtgct gggattacag gtgtgagcca ccgtgcccgg cctgcacact gctttttaat 16560

aagctgatct gtcttcttgt ttacataggc tgtgctatag tcatttaaag ttcaaaatta 16620

ctgctaaaaa gtaattaatc ctttactgaa atattcctgc tatttcatga gctgtcttac 16680

aatttggagg aatctatttg tcactgatgt ttaaaaaaga aagtcacagc agggaggctt 16740

tagcttcatt gaggaacatt attataaaat aaaatgatcg agggtaattt ttttttgtcc 16800

ttgtggtact gtttctgcag atctacatta atctgagcat ttaaagtaat aataaatttg 16860

ttatatcctc tttattattc acttcaccat ggtagcccct agtggtaaaa ggagatgtaa 16920

attgctcagt gctcacattg cagggaattt tttcctcagt tgaataaaaa ggagttaagc 16980

ctctttggaa atgagggcta tagcctgtca ataactacat atattcctgt ttttaagaaa 17040

tcagaattct ggtgtccagt aaatgtagat gccagcatta ttaaaaagtg attgagcaga 17100

gaaatgaatt gaatttttga cccactctgg cttccagctt tcatgggagc ctatacaggg 17160

ggatgatgaa gagggatata agagaataaa tgattgctct attaatctac tttaattgat 17220

actaaaatct gacagcgaat tacaagttcc aggcagtaag gcttaccaat tcattggtaa 17280

tttcaaggtt ctactttgct ttctttttgc cttcagctcc ttgcaaagga cacctttttt 17340

tttttttttt tttttgagat ggagtattgc tcttgttgcc caaattggag tgtaatggcc 17400

cttaacatcc gcctcctggg ttcaaatgat tctcctgcct cagcctccta agaagctggg 17460

attacaggca tacatcacca cgcctggcta attttgtatt tttagtagag atggggtttc 17520

tctatgttgg tcaggctggt cttgaactcc tgaccccagg tgatttgcct gcctcagcat 17580

cccccgccca aagtgctggg attacaggta ggagccacca cacctggcca aggacagcct 17640

tttatagttc agtttctcag tttatctttg actattctta attaagcttg acctggttct 17700

ccaccactga cgctattgga gttgcaaagc attttgctgt tataatgata gctcccttga 17760

tggctgttca ttattaactg gttttcaaaa gtgctgcttt taactttcat tttataaata 17820

tttaactcaa tttttataaa atgtgcatag actatagagg ttctgcttac ctgtttaata 17880

cagtaaaaaa atcccttttg taatcttata taatattaag aatgtgtcct taattcatgc 17940

ttaacataaa ttcttacaca actagcagta tctcaaaaaa taaatgattt aatagtttat 18000

gacccttata catataatta attgtagtaa aatatcttag aaaatagaat gagtattttt 18060

tataaagata aaaaatcctg attttttttt ctggtcttaa tataaaatta aatgtgttaa 18120

agggaactta taaattacat agtagttcat tgctcaggaa aagaaatgac aaagtaatta 18180

agagtgtaaa attaaaagga ttaatttttc ttaaaggccg atgagttaac aacacgcatc 18240

aatctctctt ccttctccaa acttcagaca aatgatggca aagtaatttt taagaagtca 18300

atttatagag gcaaaaaacc tgaagtacag agagtaggaa agacagtaat agaattgaga 18360

gcagttgaca tccataacat tcctattatt acctccattt tctggataag aatatggaga 18420

agaggaatcg ttaagcaatt tgctgattgt cacatattca gtaagtggca aagttagtat 18480

tgaaacccaa gtagtcttgt tttaaagtca tacactcaat cactgtgcca taattcaata 18540

aattgtgaag gatgggaagc agatggagaa gtgattgatg aagcaggact gagaaagtgc 18600

tgcctatgtg cccacagtgt gaagacccta caagacatca gaacatcatc cagtgaagct 18660

gtgagcagct caccacttgg aaaaactgag tgttttggaa ggcagggata aagtgcagtt 18720

ctgaaaagag aaaaatccat caagttctaa acggtcgcgc tctcttaccc cacagtgggc 18780

aatggtctct ccattcatca ttttcttttc cctgagtccc ttgaatctta aatatactat 18840

tctgtgaatt aaacatggta aacattagac tttgaccagc gctattatca gaagttcaca 18900

tacctaatca aaggacctgt aaccttcttc ccctccttag tttctacacc accagtagcc 18960

aattatacta tcccagctcc catcctaaaa tccccttcta aaatagaagg caaaagatca 19020

tcagtcattt gagcaaaatt tcagcatgac agcatgagta gttgagacct atagtaacaa 19080

ctaaaatgga ctgggataaa acaaataatg tttggaatca aagaacagga aaaatacaac 19140

cgatattgga gcaagaaaaa ataaaaaaga atatgaaaaa tagataagaa atagctattg 19200

gaaaggtaca agaaatacaa ttttaaaacc agtggaacat aagacaaacc acaaaatgag 19260

aaaattctat taataaaagg tcgggggtcg gggaactgta ttcttttttt tttttttttt 19320

ttttttaaga cagagtttcg ttcttgttgc ccaggctgga gttgcaatgg cgcaatctcg 19380

gctcactggg caacctccac cttctgggtt taagctattc tcctgcctca gcctcccaaa 19440

tagctgggat tacaggcatg cgccaccacg cctggctaat ttttgtattt ttagtagaga 19500

ttgggtttca ccatgttgat caggctggat ggtcttgaac ccctgacctc gggtgatccc 19560

aaagtgcagg gattatgggt gtgagccacc gtgcccggcc actgtatctt ttaaaaatgt 19620

aattgtcatg gaaggcaaga aaggtttagg aggctttcca tattgaagag gactgaggag 19680

acatggcaac taattgcaat acatgattct agactgcatg caatactgga ggaaaatcat 19740

gctataaaga attgactcaa ttgataaaat cagaatacta gcagtagatt acatcaaagt 19800

atcgtatcaa tgttacattt actgaatctg actaccatac tatagttaag taagagaata 19860

ttcctatttt taggaaatac agaagtgttt agaggtaaag ggtcatgata tatacaactt 19920

aacaaaagac ttagacaaat atggggtatt taaataaagg agaatgacgt gcaatctgag 19980

aacaagaagt ccaacagaga aaagcaaaaa tagggagtca gcgagcaacc agtgagatgg 20040

gagcagaagg atggaagggt ggaggagtga catctaaaca tattaagaac tcaaatatgt 20100

tgattatttt gtatatctga agtttgaaat tatcactgac aatcttctaa acttcaggaa 20160

aaatgaggat actgtgctac aaatgtaaac ttaagaacaa tatttacata gtcatcataa 20220

tgtaattatg aaatactgat ttcaccaaaa ccaatttgat ttaacacatc gagagaacag 20280

tgggaataga gtcctgtaat tggtgcacct gtcctaattc ctatgtaggt attataatag 20340

aaagttaata ggtaatgttt aaaattgata aatcaagata acaatatcag catgttaggt 20400

taaaaacaag gtaattacca aaagcaccag ataaaagagc caaacattgc tttctctatt 20460

gaacgggact acagcataga gaagtgttaa gcaagctgcc attattcttt tgctcttatt 20520

ataagcctct taatactatt tgattactag ctcatgagct cctatttata aaaataaaag 20580

caaattttcg aagtcagaac aattcatgct gctacaaaat ccctaaagtc atttcaacaa 20640

tacagcacac tgatgattat atagacattt ttcttgtctg agttctttgt ctagaaactt 20700

tttaaaagaa acgtggtcct gtattacatg tagcatgctt tggaagaaaa actatctcca 20760

atcaaaaagt ttttaaaaga gcccagatgt tatcgtcctt tgtatctcta aaagcaattg 20820

cttttataaa attaggatct actctttttt gaaatttgaa tgctgaagtt ttgtgggttt 20880

cagcaaaaga taggcataac atacatctct acattttatt aaatatgatg gtgcagatgt 20940

taaaaatatg tttgaaatgc agtaccttct gttcttgctt tcaactccta ccctagagta 21000

aatgaaatgc taatcacctc ttgtgttagt gatagaattt gtgagtttcc acagataaaa 21060

tatgtttact gtaatttcca aatagttgat agccagtagc aatatcttga cttcagcaac 21120

tggtatgtaa gggtctatat taaagtgttt atattatgac tagtataaca gatggtatta 21180

tttgttacag caggtcataa cactaaaaaa taaaactatt acacatctac aatttgatca 21240

caaattttag tggcagtgat cacttatttt atgaaagaga ggcagtaatt gccgaagtac 21300

tgcctaaaaa tgtaagacaa ttctcattac agggtaaggt aaaattattg acttattaac 21360

cagaaattca catctatccc gctacaggaa aaaaagtgta catttaatat ctactacagt 21420

cagttgagtt ttcaaagtaa tctgtcatct ctaccatgaa tatggggaac aggagaataa 21480

ctgctggatt tttggcattt gacacctaat atatagaact ttaaaggagg ttatctatta 21540

tataaataat ctttgaagaa gagcaattta tgtttttgga caaccaaaaa attacttgtt 21600

tagcttcaca ggaattttaa agcatgtttt cacttattat taaacagtaa cattgaatat 21660

actgctcttt tatcagttct catttttttc aacattcagt agatgacttt aaagaactct 21720

caccttcctg gttttctttc taaactgaga ttatttcttc tctcacttat gcacctattt 21780

tctaatggcc atgtctcaaa aaatttttat tctagcgtct tttttccttt tatcttggtt 21840

ttgctgtagt gctagataga tggttacttg tgagactttc aaatctatat accaggctcc 21900

aacttctctc cctctccctc tctttttgag taagggtttg gctccatcac ccaggctgga 21960

aggtcatggc tgattgcagc tttggactcc tgggctcaag ggatcctccc acctcagctt 22020

cctgattagc tgggactcca ggtgtacagc atcatgacta gcaattccat tgtttttaga 22080

aatggggtct tgctctgtcg accaggctgg tctagaactc ccagcctcaa gcaatcctcc 22140

tgcctcagcc tcctaacgtg ctaagattac aggcgtgagc caccacacct ggccaacttc 22200

taagtttcag tgctgtagtt ttaaattgtt tctcatactt atctaatatt ttttcccacc 22260

atgtcaaacc cttaaaaact cagtatggac aaaataaata ttaacttttt tgagattgat 22320

aattgtatga tagtgcttat tacctcaaaa gaattactgg ggcagcagta taatccagta 22380

gatgggggaa atagtggata ttaagaccga aatgggatca taacttttta gttgtatttg 22440

cccacattaa ttgttttaaa gggggtaaag gcctggaata tgaaaacagg aggatataaa 22500

atatatggag gttgtgaaag atgactcaga acttgaaaca attgtttcac tgcagtaaat 22560

acatgttgaa tgcatagaac agagaaaaac agaagaaaag aagaaggaaa aacaaactcc 22620

atgatttaaa atattgatga atgtgacaat aattgtatct ttaaaataac tgagaaatgg 22680

tagaaataaa aattataagc ttcaactgta tattacttat tcttgacaaa gatgatagtt 22740

gcaagctgaa gccaaaattt ctaactagga ctaatgcaaa gatttcaaga gaaactttaa 22800

taaagaatta ttgatgcaag cgctttcaga aagcaaaggt agggactaac cattagctct 22860

tgtttagaaa gataaaggtc acaaataaaa tttgtgtttt aatctataca ttttcagcat 22920

tatgtcatca taaatttgca tgttgtttat tacaaatggg aaaaggtttc ctgtaatata 22980

tcaagatgag ttttaatagt ttcaaggacc attgatttgt aaacttaaga ctattatagt 23040

gttgtggaaa attaaatata attggtgttt taggaaaagt tgttcattga agtttccatt 23100

ttatgtgcct taacagatta attttgctta ttaattctgt gatatttaag cactgctgta 23160

agcttcacag atttggcaat agccactata tacttaatga gtacaaacta tggagtattt 23220

ccatattaag gtagaagaaa tatttaaaag cacttctact tgtgatgtca ttgggggtgc 23280

aacacaatat aaagtacgca cattgctgga ggagaagcat aaaataaaga actcacgggg 23340

cagaaactta agacacacct aaatcaaatc tttgttttca gcattatctt tcattactat 23400

tctttgttac aaaaaaacat tgctatctgt ccttttctat atgttctcta cattaaaact 23460

tattatttgt atacatactg ttccctacat ctgcagtgcc attcccagct cacatatctg 23520

ccctcttaaa tttatatcaa attcaactcc cttcaaaaag ccttctcagt tgctttaacc 23580

caagccagta gtctgtcttc tagaaaacag ttgatgccga tgcctccttt tcctttagca 23640

actgaaaata atattatagt tatttaatgt gtgtgtgtgt gtgtgtgtgt gcataacatc 23700

atactattta gtagtatcta tatctatcaa gtatctgcta cagtggatga tagatgactt 23760

aaagacagaa atatgttatt catatctact tctcacaggt ctagtgctgt tgcaggatat 23820

tttttaagga atcagagaga ctgatggggt tcaggaggat atttattatt taggtgcacc 23880

agcccagtcg gattaacatc caaaggactg agccctgaac aaagagttaa gttacctttt 23940

aagcattttg tagggtggga ggagatctgt gcagggggaa gcatgttaga gaagtgagaa 24000

acaaagacag ttattcaatt aattgagaaa tgcgttacat catttcttac ttttcaagga 24060

aaaacatgtt ttgcgacttt tgtttatgtg tctagtgact ttgcagctgc ccagctaggg 24120

aatcagggtc ttcacaatgc ctgggaatgg aggggagaga aagctcatta gccacagaaa 24180

aataggccgt tagtttttta aaggactcta gctctttctc tttctcaggg ggaattggat 24240

tttcttacat acaactgagg ttctgcttac acattcttta atttctttta attcctgttc 24300

cagtgcagtg caatgttcat agtagatgct taatattttt tgaattattg tatctttagt 24360

cattgcgagc catcattttg agagtcagtc tcattctttt agccctagat tatttttatt 24420

ttcttgagat acgtgaataa agggcacagg aaaaaaacac agttgagagt tttggataat 24480

agtaacctaa ttcacccaga ttttacttca tattggttgg aactctaagt tgggtgcaac 24540

taacatgaga cttgttttga tttattgtaa acatattctt gaatgactaa tgaacagtgc 24600

ttcatgtgaa caaattttca ccttctgata atttccaata ttacactttg aaaaacattc 24660

ccattgtagt gctctagaaa atataatttc aattgttaaa attcaatatt actgtaattg 24720

aggatattaa ataacttgtt gttacatgat gctagctgca gcttaaattt cagatcatga 24780

gtgttgttaa tattatatag ttggaattag ttctgagatc aatgaatcaa tgctgtctta 24840

ctgttttgtc tttaattttt ataccacatt tattttgtga gtggatttca taaaaggaag 24900

ttgctttatg ttgccaagct cattatcttg ggtttttgtc cagagctgaa ttcccttttt 24960

catcaaagga cagagagagc ccattataaa ctattgagta atgctgattt gatgggtagg 25020

ccacttacct ttaagtggaa aggaggccag gcgcggtggc tcacgcctgt aatcccagca 25080

ctttgggatg ctgaggaggg cggatcatga ggtcaggaga tcgagaccag tctggccaac 25140

ttggagaaac cctgtctcta ctaaaaatac aaaaattagc cgggcgtggt ggcgggtgcc 25200

tgtagtccca gctactcggg aggccgaggc aggagaatgg cgtgaacctg ggaggcggag 25260

cttgcaatga gtcaagatca caccgctgaa ctccagcctg ggtgacagag cgagactcct 25320

tctcaaaaaa aaattacaca aaaatgttag actaataaat attaataaac actaaaggaa 25380

gaaatttagt ggaacattta tcagaggcaa cagaccagga agttatagtt accctgtgaa 25440

aaggcatcat ctaggattca tagaaaacat ttgcaaattg gcaacaacca ccaccacaac 25500

aaactaaaac aaatgaaaga aatagacatt tatacaggga aaatataaat agataataag 25560

tgtaataaat tttgtgactt tccacttgaa aacaaagaaa tggaaataaa acctatgtaa 25620

agaagtggtg gagtgagccg agctggggtg ggaggggtgg gggcatccca ggcagaggga 25680

acagcagatg caaaggtctg gaggctggag caggctgtga ggggtgagag cagcaagtac 25740

atgaagtcta gaccacaaat cgatgcttca ccctcaaaaa tagcctctca ctggccacca 25800

ctcaggccca gatacatgtg taaggtgggc agccttcagg aaaagtgacc gcaaaagtgc 25860

accacattgg gaattctgag gtgtttgaga aagttatgga tgaagggtga ggtagactct 25920

agtggggaat gtcctcttgc cacttggact tctctgtcta tttcatgggc tattatcaac 25980

ttctaaagag ccagtggttt gggtgcacat ttaggtgtaa gtacctcttg cctcaagcct 26040

taactctctt tccaatgagt caagtgagct gttgtgttta agtaacttct ctttattgtg 26100

aaatctttcc tatcacttta agaaccttct tcttcagacc caagtaatcc caaggcacct 26160

ataatgtccc tcctggagaa gacaaaaggg tgtcttctat ttctgtgtgg cccgatattt 26220

cagggcccct aaccaggtta gttcaaggtg ctcagcataa ccatgaatct gctacccatg 26280

tttctcaggt gggtcatagg ccctggaggg attgtggctg tggaccaagc agggggtgct 26340

gtgggctgtt ctgagaagga gggttttgga gtgtccaagc tgagaacata gaggccccag 26400

atgttgtatg agtaaatttt catctgacag aattatattt tcctaactga aaacatgaaa 26460

cagagatgta acaactgtgc ctctttggct ctgtctttgc atgctgtcat cagccactga 26520

acaggagctg agagggaaga cataaagaac agaaaaccca tgagggggat ggtccatgtg 26580

ggtgtggaag tgagggtgag tggggatttg gccaaagagg cctaatccac aggagtagag 26640

tctcatgtgt gactttcaga atgagcagga gtgagatagg tggaggggga gtgttggaac 26700

atctcatcag aggtgagaat ccatagaagg agacactggt cattacagtc agtgagaaaa 26760

gtgactttag cctgagccta aggtcattgg atcaaggaag gggccaggag gagctccttg 26820

aagggaagct gtgagcttga gaggcaggct gaggcagatc agtagaatga gcccccgaga 26880

agggctcaca gagtcactga aggactgtga ggtggcaggg acaagctcac atggcctttg 26940

gtcagtgagt ctccagctgt aatgagaatg aaatggggaa gatctgggac aggaagacaa 27000

gctgggggcg ctggtaccat catggggaga tgatggtggg tggctctagg ttagttagag 27060

ttggaatgaa gagcagaaag cagcttcgtg atacctttat gtcataaaat acatttggca 27120

tggagaggac ttggactttg tgcatgaagg atatggatgt ttctaggctg actcccaaat 27180

gtttctaggc tgactcccag aaactaggat gtttctaggc tgttcctggc tgagaatttg 27240

gatggtaact atagtccaaa gaaacaaaga acactgaaat catatcttat tttttttaac 27300

tcacaaaata gaggcaggta gatatttata tgtgttttgt tggctcaatg atttttttta 27360

gtaataaaca tatttactta gaaactactg ggtgtctgta tgctctggga tttttttctc 27420

ctgtcctgat gacatctctt tcctttctct gggttcagga gttacttccc aggttgtggt 27480

gacacaggaa cattcacttt ccacaactcc tggaggggca gtcaccttca cctatggctc 27540

caaaattggg gctgtcatca gagcatctcg acagctgggc ccaacagaac atctggcaat 27600

gtccccaggg caatggtggg cacaagcagt caggtcctgg gacccctgca caatcccctg 27660

gttctcttca tgaagaaaaa gccactctgg ttataatgag gactagacca agaatgaggc 27720

ccagtgttac atgctcggtg tggtgatctg aaatatcacc acagggacaa atacaaaaat 27780

acaagtggga aactacactc aaacccttgg atcaagcctc agtcctatgc ttctgaagca 27840

aatttcttgt ttttggagtc tggttttcaa ccactgtaga ctcagatcat ttattcttca 27900

ggcccccaat gggatcatca caagcacttt ttctatcaat catggcagtc acttgagatc 27960

aaaggtgaat ttcttcttcc tcaagttata agcaacactt tggcatattt ggtgaacttc 28020

aaagaaaatg gctaagatag aggatgagag acataagagg cttcaggaat atttcagcag 28080

atgcaaagca gactacaggg taactcattt cccatatgat ttgattgatt gcaaaatgtg 28140

aaattaaatg catacaaagg ggaatgtgtg tcatttttcc tcttaatgac aaagtaaaaa 28200

aaaagcaata aaacaaaaac aagcactatt tcatgtgtct tgataaactt gtagaaagtc 28260

ttataatatc cacaatatct aggatacagt aaaataactt tccatatgaa gaaccagaag 28320

aacacaactt aaatgagaaa agacaattga tagacaccaa tactgaaata atttaggtgc 28380

tgatgttatg taactgagat taattaagga gtaattatac aattccttca gtgagcaatt 28440

ttagacatta ttttttgaag ttttaggttc aggggttcat gtgcaggtat gttacgtagg 28500

taaacttgtg tcagagggga cattcttgaa acaagtggaa aagaagcaaa tcccgaaaag 28560

aaaaataagt catagaaaca aaccaaatta aaattagaaa acagaaaaat gcaataaatt 28620

aaaaagaaca gtctggaggg actctggaac aggtaacagt gaaattaaag ttcaggcatt 28680

agaggaatta tttaccgtta agtagggaag aaagtacaaa caatagccat ctaagcccag 28740

cagagggagc tgtggatctt ctcagaggtg agcaaggccg gaaatgaact cccggggctc 28800

tcatttatgg ctcctgggtc catactcact ggcccgaatt ggggtttaga gcaggtgctt 28860

tctcccaaag ggcaaactca gagttcccag cagcccatcc tccctgacag agcggctgct 28920

gcctggctga gcactcagaa cttagggagc cggtggtttt ggtagaggcc ccatgtttgc 28980

ataatgtcct aaccaaacac acatctaccc ggggaggtga cgaggagtta gaggaactgt 29040

cccagggtcc agaagtagcc gggggttctg gggatccttg gaaagtccac accatagctt 29100

ggatgtctat cctcccctct ctccctcctc actgcccagg ttcatgtaga tttcaaagac 29160

aggactttgg agggatctgt ctgctgtgat tcctcacaca taacatgtat ctgttttagt 29220

ttcttggttc agttctcagg ctgtggtgac tcaggagccc tcactgactg tgtcctaaag 29280

agagagactt attctcatgt gtggcttcag caccacagca ctgaattcag cagaagcctg 29340

gtcaagtcct caggacattg atgtgtaaca cagataacaa acacccctgg atgcccactt 29400

gattctctgg ctccctcctg ggggaaagct gccctgaccc ctcaaggggc ccagcctgag 29460

gatgaggctg agtatactgt gggctacacc acagtggtcc ttagcacagt gagagaccca 29520

gatggggaag caggacatga acgagctttt ggctgatccg gggtacaaac atgaacgtag 29580

gatatcagga tcacgtggcc agacacctga gctctcaagg acacctaaat tttaactatc 29640

cgtagagaga aaggtgggtg gaggggtgta aattgattcc tgtatctgtt tgaattttaa 29700

agtggaaaat atttgtgaat catttgggag atgcagattc ccaaaaagaa gaatttatgt 29760

ttatattcca aacagaacag taatagtgag atcaactcta aagactaagg tcaaacacac 29820

ttcatccaga aagatggatc tcaaaagatt ttgttgttgt tttgcctatt aaaagctaag 29880

agaggaaatt atattaataa agtttatttc atgttctatg ttaatagtgt gtatttcata 29940

gctttttaag cattaaaaat tataaaagtg ttaagaggga ttctcttctc taatgttata 30000

taagaaagta tccacctagg aaaatatttt gaaggtctaa cccccagtac ttctaaatgt 30060

gaccttattt ttaaaatagt gttattgctt atgtaattct taggatgaag ccatcctaga 30120

gcgaggtggc ctcttgatcc aatgtgcctg gtgtccttat aagatgatgg tagtaaggag 30180

attggaagac acagagagaa tgccgtgtga cgacgaaggc agagattgaa tttacacagc 30240

tgcaagataa ggagcactaa agattgctgg caaaccacca gcagccagaa agagggaagg 30300

aaggatctcc cctcaggtat atatggaaac attgcccttt ctatgaattt tgggaggcta 30360

gcctccagaa ctgtgagaca atacatttgt gttatcttaa ccaaacttgt gtttaataca 30420

ttctgacagc agttctagga aaccaataca ccagacaata aattacacaa tacttctgaa 30480

attgattgta cttctttcat tttggcaacc agtttttcca tgaatatgat tatatctgtg 30540

ctcatatttc taactttata aatatgcttg taatttaagg ttaagataag aaatccttgt 30600

gtcttaaaaa tccctaaata tatgagaact gaataataat attaacatta tgagatctaa 30660

tggaaaattg tagcttttac ataattccac ttgttcacta gtatcaagat gtgaaggtgg 30720

gaagatcacc ttgttattaa caaaacactc ctggtcatct ctgtggattt gagagtgcag 30780

ggggagacac ctacatattg tttgggaagt cacaaatgca gcatgcccag agcatcatgt 30840

acaacagtgt gagtttgctg gacatgtggg caggacgcag aacagactgg ggcatgagga 30900

accacctatc accaaaccca ccagcgcctt cctccctgga tcagcccaga gctgctcaga 30960

cttgaatgtg cacatggatc atgtgggatc ctggataatg tggattctga tcagtgggtc 31020

tggactgtgg actgaaattc tgcgtttcta gaaggcccct tcttccaaag atccacatac 31080

atgttttaat attaccatat tcataagaat ggtgggaaat aagggaagcc actttcagac 31140

agaaaccaaa tagaaattta tctgagaggg agctaaagga ttatgatcaa agagaaatcc 31200

ccaattccag gcagggagct gatcacagag ggcagatggg aagttgggag aggttctatt 31260

tttttttttt taagaaatag aaatgaatgc aaacacaact caaacctcct cttgatccaa 31320

catccctgct ggctcctgcc caatttctgc actggcttct tctccagtct tttcccaagc 31380

atggaccctc ctcaccctct ctgcttcccc atctcaaact cacaacttcc cttctcagtt 31440

ctcctgtttc tctccctttc attccacttc cttcttctct gtgacctcct tagacaatca 31500

ccaaaatatt tctagaagaa aagccatggg tatttttcat cttttcttga aagttgtcac 31560

tcacattccc tcttcgttct tcagcctctc ttacccctta ctacaaaaga tgcctgaggc 31620

cactgtttct cttttccatc ccctcaaata gcttctcctt ccaacccgtt tctctcctca 31680

gttactagag cattcagtga agcactcagt caggttaaag gatcctccca aatgctcacc 31740

ctgctctcta tgtgtgatct cctgaagaat caagtaggga gagaaacatg gcttcatgag 31800

tttgtgggga agtgggtgca tggctgaact gtccttgttc tcagcaaatg tactttattt 31860

catgcacagt ttgtatatga gacaaccttc tcctgcggac tccttataaa atgggaagaa 31920

ttatgtgttg tgttttcaca gaggaagccc aaatattcca tgctcatggg taggaagaat 31980

caatatcatg aaaatggcca tactgcccaa ggtaatttat agattcaatg ccatccccat 32040

caagctacca atgactttct tcacagaatt ggaaaaaact attttaaagt tcatatggaa 32100

ccaaaaaaga gcccgcatcg ccaagtcaat cctaagccaa aagaacaaag ctggaggcat 32160

cacactacct gacttcaaac tatactacaa ggctacagta accaaaacag tatggtactg 32220

gtaccaaaac 32230

<210> 75

<211> 117

<212> PRT

<213> Intelligent

<400> 75

Met Ala Trp Ala Pro Leu Leu Leu Thr Leu Leu Ala His Cys Thr Gly

1 5 10 15

Ser Trp Ala Asn Phe Met Leu Thr Gln Pro His Ser Val Ser Glu Ser

20 25 30

Pro Gly Lys Thr Val Thr Ile Ser Cys Thr Gly Ser Ser Gly Ser Ile

35 40 45

Ala Ser Asn Tyr Val Gln Trp Tyr Gln Gln Arg Pro Gly Ser Ala Pro

50 55 60

Thr Thr Val Ile Tyr Glu Asp Asn Gln Arg Pro Ser Gly Val Pro Asp

65 70 75 80

Arg Phe Ser Gly Ser Ile Asp Ser Ser Ser Asn Ser Ala Ser Leu Thr

85 90 95

Ile Ser Gly Leu Lys Thr Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser

100 105 110

Tyr Asp Ser Ser Asn

115

<210> 76

<211> 32230

<212> DNA

<213> Intelligent

<400> 76

gaattcactc atcaccaagg ggaagatgct caatcattca tgagggatct gcccccttga 60

tctaatcacc tcctagtggc cccacttcca acactgggaa tcatattaca acatgagatt 120

tggaggggac aaatatataa accatatcct tccacccctg gtcccccaaa tctcatgtcc 180

ttctcatgtt gcaaaataca atcatgcctt ctcaacagtc ccccaaagtc ttacctcatt 240

ccagaatcaa caaaaaattc ccaagtccca agtctcatct gaagatgagt cccttccacc 300

tatcagccta ggaaatcaaa gacaaattga ctcccaaaat acaatgagag tgcagacatt 360

gggtaaacat tcccattcca caagggagaa gttggcaaaa agaaaggatc tacatgcccc 420

atacacgtca gaaatccagc agggcagtta ttaaatctta aagctccaag gtaatctcct 480

ttgacttcat gtcccatatc cagggcacac tggtgcaggg ggtgggctcc caaagccttg 540

gacaactcta cccctgtggc tttggaggat gtcgcccctg tggcttctct cacacgatag 600

agttgagtgc ctgtggcttt tccagattca gggtgcaaac tgccagtgga tctaccattc 660

tggggtctgg aggacagtgg cccccttccc acagctccac taggcagtgc cctggtgggg 720

acattatatg ggggttcaac cccacattct ctttggcact gctctagtag aggttctctg 780

tgagggctcc atgctggcag gaggcttctc tctgggcacc tagggtttct catacatttc 840

tgaaatctag agtgaagatg ccaagcctcc ttcactcttg cattctgggg gcctacaggc 900

ttaacaccag atggaagttc ccaaggctta tagtggcttg cactctccaa agaagcagcc 960

taagctgtac ttggggccct ttgagccaag gctggagcca gaggaaccag gatgtggaga 1020

gcagtgtccc aaggctgcac aggcagcagt ggccatgggc atggcccaca aaaccatcct 1080

ttccctcttg gcctctgggc ctgttatggg aaagaccacc tcagagattt ccgaaaggcc 1140

ttcaaggcct ttttcccttt gttctggata tgagcactta gctccctttt agttatgcta 1200

atctctctag caactggctg ctccatagcc tacttatatt tgtctcctga aaataccttt 1260

tcttttctac cacatggcca ggctgtgaat tttccacatt ttttatgctc tgcttccctt 1320

tgttgatgaa aaaagccaaa ctccataaaa cacttgacga gattgattct gagccacatg 1380

tgagagccat gaactgtggc acagcctcaa aaggtcctga gaacacctgt gcaaggtggt 1440

tgggttgcgg cctgctttta tgttttaggg agacatatca atcaatacat gtaaagtata 1500

cattggtttg gtttggtttg gaaaagcagg acaattcaaa gtggggaatt ccaaatcata 1560

ggtggatttg aagattttct gattggcaat tggctgaaaa agttaaatta tctaaaaagt 1620

tgaagtcagc aaaaagcaat gcttaagata agggggttgt ggaagccaag tttcttgtta 1680

tgtagatgaa gcctccacgt tccagagaga agagatggtc aatgtctctt atcagaacct 1740

aaaaggtgcc agactcttgg ataaatctct cctgaatcag gaagagacct ggaaagggaa 1800

aaagattctc aacagaatat acatttcctc cacaagagac ggctttgcag ggcccttcca 1860

aaatgtgtca gagaaatata ttctggcata aaatacttta atttccttcc atgcctgcca 1920

cctgtcatgt gatgctatac cagaatcagg ttgggatttg atatcttatt gctacaaaga 1980

gtctgctttg tcagtcttaa gctccctgtt ttaatgttaa cactggtcag ctgagcctaa 2040

gctccaacag ggagagggta tagtgaggga gtccaaacca ccctcccctt cctgtcatgg 2100

cctgaactgg ttttccaggt ttctttggaa tccccttgga ttagcatttt attttgtcgt 2160

ttacaccttt taaatataag ttccaacttt aagtcatttc tttgctccca tatcttacca 2220

tagactgtta gaagcaacca ggtcacacct tagaaacatc tgcttagaaa tgtcttccac 2280

agatgcccta agtcatcact cttaaggtca accttccaca gatccctagg gcatgaacac 2340

aatgcagcca agttgtttct taggatttaa caagcgtgac ctttacacca gttcccaata 2400

agtttctcat ttccatctga gatttcatca gcctggcttt cacagtccat atgtctatca 2460

gcattttggt gacaaccatt taatagtctc taagaaattc caaactttcc ctcatcttcc 2520

tgtcttcttc tgagtcctcc aaattcttct aacctctctg cctgttacct gattccaaag 2580

ctgcttccac attttcaggt atccttatag caatgttcca ctcctcaata ccaatgttct 2640

gagttagtct atttgtgtta ctagaaaaaa aaaaaaaaca tgaatctggt aatttataaa 2700

gaaaagaggt ttgcaccagg cactgtggca aggaagggag gattaactag ctcacagttc 2760

tgcagccatt ccggaaacac ggccctggca tctgcttctg gcgaggcctc agggagattg 2820

caatcacggc agaaggcgaa gggggagcag gcatgtcaca tggcaagagg gagcaagaga 2880

agagaagtgg cacgtcctag acttttaaac aaccagctgt cacagggact aactgaggga 2940

gaactcactc atcacctggg agatggtgct aggccattca tgaggggtcc agccccatga 3000

tctaatcacc tcccaccagg cccacctcca acactgggaa tcacatctca acatgaaatt 3060

tggagagaca cacatccaaa ctattgcacc acaggaagct ggaagaggtg gacggtcctc 3120

ccctggagca ctcagagaca gccaccaaca ccttgatctc agattcctga ctcccagaac 3180

tctgaaaaaa tttttgttgt tttaagccac atttgtgata ttttgttacg gcagccacag 3240

cacggatata gctggtaaca gagttcttta gcctccaaaa tacaagcctg gggtaaataa 3300

ctattgaatt tggccacatg gaaatcagga agaatgtaat aagcatcatc gatggacggg 3360

atgacctggg agatcgtggc aagtttaaag ctaattggaa gttggcaacc aaaaaatcct 3420

gtcttgcaaa ttgccaggaa attgccaaga aaacttttag aaaacatggt taaagatgga 3480

gaaacctgga tgaatccctg gtttctgaag aggtgcttag gcattgaagt gagactgatg 3540

agacttcaca ctgtaccttc agaagggcag acactcacga gatcatcaac cacggctccc 3600

tcgctgggag gcagacacag agctggaggc ggcggccaca aactaggtgg gcagagaggc 3660

gtcagcatcg ggacttgcgg gaggccctgg gaaggaacag gcagcggtgg ggcctggaca 3720

gccctgggga aaggaagtta cttggaggag accaggggct tgcatttggg ccaaaaccaa 3780

ggacaagaga tcagctggtc ccagggaact ctggagccca gcaccccacc acgggtgcag 3840

ggtcctgaca acccgagagg gctgtgggca ccgggcacca gcctgtgggg attctgaagg 3900

ctgctgccca gccacacgtc ccggttctga acaaagggtt caagagacac ctgcaaacct 3960

acctttctga gagcagcctc ttagcctcag ctggtattgg cctttggtca ccccacatgc 4020

cccgacagtg gggcctggcc tcagaaaggg gcccctccat ttgtactttc tatctgatcc 4080

ttgacacagt gctaacacca aagaccaaac ccgaatcttg gttcacatac tctgcaaaga 4140

aatgactaag actagttctt ccggaggtca gttttagaac gtttattctg acttaattct 4200

gccttatctc tgtgcaccaa gaaaactatc tgcacatctg tagtaaaatg tgaaggtacc 4260

ttcattttgt ttgtatacta aacttttaag aaatacctca ctttttaaaa cttggttttg 4320

tgttttgtat gcttataata ttttccataa tagatcattt taatgatgaa ctctgagtct 4380

tctgacagaa tgcagatcaa tggattccta gccatcaggg aggcgttgat tgcaaagggg 4440

cctggtggaa tttttggaga gattctacat cttcttgcaa ctgttcatct ggattacggt 4500

ggtggttgca tgactgtgca tttgttgaaa cttacatgtc acaggatgga ttatactgaa 4560

tgtaaactac acttgagtaa atatgacttt taaaacttta aaaagttggg ggatggttca 4620

ctgccccggc tcgaagcccc ctggccacgc tgcctggcca gcccaccccc atccctgcca 4680

gcgcttgcct ccagcgtccc aacagcctcc tgaccccctg ggctgctcta aaccctcagg 4740

agcgcagcca tccgggatca gctggatgga gatggggagc ccgagactcg tgccacacca 4800

cgtcctcccg cccacaccag ccacacgcag acgtcaaagc agcactgtcc tcgcacgcac 4860

tgctcccacc caccctggca cggccattcc aggcctgggg caggaaggca gatgctcccc 4920

ctgcccccag acacaagcat tcctgcacac acccccagca cacacacgca ctcccatgcg 4980

cacactgaca cacacaggtg tgtgcagctg agacacagcc cgttcccagg aagcccagcc 5040

cccatcactg agggaaaggg gcagcaccgt agggccacag gagtggcagc tggacacaga 5100

gccaggagct ggtgaaggcc caggccactg aggcgggctg ccggcatggc tgggcgtgaa 5160

ggccagaaga gggcaggagg ggctgggggc actatgccta ttgggcctag gtgggcacac 5220

gccgggcagg agaggaacag cccagcccct cagacaggaa ggggtggggg caggggccat 5280

ttgtggaggc cagggcaggg ccagcacccc aaggaaaagc agagcagggt gagaacggac 5340

atggggctca gagctgagca ggcctgctgg gccccaggag ggagacacag atgaccggag 5400

atctcaaggc tggcagaggc cagagatgga gccccagctg ggaagccatc ctccttcctg 5460

ggggcccacg ctgcccggcc cctccagccc agcaagcttg gggcattgga tagaaccggg 5520

agagagccga ccaggcactg aggcccctgc cccaaatgcc cacagcctgg ggaaaatgag 5580

caggtacatg ggaggggcaa gtggagcccc aggcacaccc acacagtgca cacggcctca 5640

cctgggccgg agggggcagg aggctcgcca ccccgctgtg gtttctctcc taatctcacc 5700

ctgggtttct ctcacacttg atgcagatga tgtttctctg acattgtgga ctaagagttg 5760

gtgctggaag gggttagcca tcttggagat gttgctatgg gatgcaggga ttttgcgtgt 5820

gagaaggaca tgattatggg gggaacggag ggcaaactgt catgggttaa aatgtgtccc 5880

ctataaattc atgtgttgaa gtcctaaccc ccaggaccgc agaatgtgac cttgtctgga 5940

aacagtcttt gcagctgcaa tcaagttcag atgaggtcac cctggagtag ggcaagcctc 6000

tgatccaata tgactgctgt cctcatgaaa agggggaatc tgggcacaga cagcacgtgg 6060

ggagaacgcc ctgtgaagat ggtgctgctt ccataagcca agagcaccag agacggccgg 6120

caaagcccag cagcaaggag agagcctggg acagagtctc ccatgacaca gaggagccag 6180

ccccaccgag gcctccatcc cagatgcccg gcctccagaa ccaggacgga ataaacgtct 6240

gttgtttaag ccacgcagtc tggggtgcag tgttgccagg gccacagtta acggatacga 6300

gtgttgtcct gagctgccag ccccacaggc tgcacgaggc ctccctgccc cagcccagtg 6360

cagactcccc agccccctgg gtgtgccatg ggcagtgggg ggcccctcac tccgtcctcc 6420

cccagcctgg gaggttgagc ccattatgag ctccatgggg tgaagctgga acgagaggct 6480

gggagccgac tgggagcctg cggctggagg atggatttcc ccagggaccc acacgtgcac 6540

ctccacctgt ctcctggaca ttctctctga gggcagggct ggtgccagct cagggatcca 6600

gcagggtcac aagggcaggc cgggtccttg tggagagcac atttagtggg agggacatga 6660

tttcccttca aagtgcccat tctggacgct tcccgttcca tgctggacgc ttcctcttcc 6720

acgctggatg cttcctgttc cacgctggat gcttcctgtt ccacgctgga tgtttcctgt 6780

tacactctgg atgcttcctg ttccacactg gatgcttcct gttccatcct ggatgcttcc 6840

tgttccatgc tggacatttc ctgttccact ctggatgctc cctgttccat gctggatgct 6900

tcctgttcca tgctggatgc ttcctgttcc atgctggaca tttcctgttc cactctgcat 6960

gcttcctgtt ccactctgga tgcttcctgt tccacactgg acgcttcctg ctccacgctg 7020

gacgcttcct gttccatgct ggatgcttcc cgttacattc tggatgcttc ccgttccatg 7080

ctggacgctt cctgttccac gctggacgtt tcttgttcca ctctggatgc ttcctgttcc 7140

acgctggatg cttccttttc cacgctggac acttcctgtt ccgcgctgga cacttcctgc 7200

tccacactgg acgcttcctg ctccaggctg gacgcttcct gttccatgct ggatgcttcc 7260

tgttacattc tggatgcttc ccgttccatg ctggacgctt cctgttccac gctggacgtt 7320

tcttgttcca ctctggatgc ttcctgttcc acgctggacg cttcccattc cactctggat 7380

gcttcctgtt ccatgctgga catttcttgt tccactctgg atgcttcctg ttccatgctg 7440

gatgcttcct gttccatgct ggatgcttcc tgttccatgc tggacgtttc ttgttccact 7500

ctggatgctt cctgttacat gctggatgct tcctgttcca tgctggacgt ttcttgttcc 7560

actctggatg cttcctgttc catgctggat gcttcctgtt acattctgga tgcttcctgt 7620

tccatgctgg acatttcctg ttccactctg gatgcttcct gttacattct tgatgcttcc 7680

tgttccatgc tggacatttc ctgttccact ctggatgctt cctgttacat tctggatgct 7740

tcctgttcca tgctggacat ttcctgttcc actctggatg cttcctgtta cattctggat 7800

gcttcctgtt ccatgctgga catttcctgt tccactctgg atgcttcctg ttacattctt 7860

gatgcttcct gttccatgct ggacatttcc tgttccactc tggatgcttc ctgttacatt 7920

ctggatgctt cctgttccac tctggacgct tcccattcca ctctggatgc ttccttttcc 7980

atgctggacc tttcttgttc cactctggat gcttcctgtt ccatgctgga tgcttccttt 8040

tccattccgg acacttccta ttccattctg gacacttcct gtgcgacacc tcctcgggct 8100

tttggtctgc ccagtccctc tggcctcata ccatcccccc ttacctccca cttccacgtt 8160

cgtccttcct cagctcctcc ctctctctag agcttcggcc tggcaaggtc cctcctgatc 8220

tcagtccagg ctcccccagc acaggtagga gacttgcacc tgcccttgga cctccccacc 8280

ctgcatgatg ccagcatccc ccaggcccca gggaggcccc atttctctct ctgcttgtag 8340

tccagtggcc ctggagtgcc actgcaactc gggtgtgccc ctcgcctctg aggaagctaa 8400

gtgccctaag ctaagcagag gccatcccct ctgctcagcc ccagggccct gccccctacc 8460

ccttcccctc acctgcacca caggctctgg ccaactctgc ccaggctctg aatgggcccc 8520

tctggctccc ctctgctgct acactgccct gcaccacctc cactcagctt cagtgtgttc 8580

atccacctgt cccacgtccc ctcggccccc aggagcacag ctggtggccc tggctcctgg 8640

cagcccatct tgttccttct ggagcaccag cctcagaagc cttcctgtgc agggtccact 8700

cggccagccc tgggaccctc ctggtctcaa gcacacacat tctccctgca gccagacctg 8760

cccctgcctg tgagctcaga cctgagcctt ggaacgcctt cccttctcca tcccagctcg 8820

cctttgccag ctgctcagcg ggatgaactc acactcccct ccctgcacca tgagtgagag 8880

ccagctggag agacgcccag gccaaagcag ccaccagggc ccagtggggg tcagaagctt 8940

caggtgagag gcccaggtat tgagaggctg agaccacggg cagaatggtc ataatcactg 9000

ccagtatcag tccagcccca gggactcaga gacagagaaa agagcagtga acaaggtccg 9060

ggctccccac cttctcccac gagtatgggg gcagccacca cccccatccc cacacaccca 9120

tgaggcagcc tcggctgtgt ctggactccc ccttaccctg tgacacagaa accaccagaa 9180

gaaaagggaa cttcaggaag taagcggtgc cgccggtttc aatcctgttc ttagtctttg 9240

cagcgtggag ttcacacccc tggggacctg agggccgagc tgtgatttcc taggaagaca 9300

aatagcagct gacggcgtgg gcaagtctgc ccacatgtac cgcgccaaaa caggaagggc 9360

tgagaccccc acctcggtga gtagggtcag cacagggcaa gggcacaggc tcgggaggag 9420

aaggacagag cctgggtgca gccgtgggcg ctcctggacc tcagctgctg aacaggctac 9480

aagaggctgg ggagacgtgg gggcaaggcc agccccacat ggagacccaa gcggagccag 9540

cacgggggag gtgggcagcc ttcaggcacc aacgcccacc cagtgcaaga tgacggggac 9600

cgtgggcagg ggcttccaag ccaacagggc aggacacacc agaggctgac tgaggcctcc 9660

atgacgacca ggctgggagc acgaggaacc tgacgggatg cggcagagcc ggccgtgggg 9720

tgatgccagc atgggcagga cccacctgag ctgaggaggc agtagaacga gggaggagga 9780

gaggccccag gtgaacggag gggcttgtcc aggccagcag catcactgga gcccagggca 9840

gggtcagcag tgctggccgt ggggccctct ctcagccagg accaaggaca gcaggtgagc 9900

cgggagcaga gcagggaggg tgagtgtggc agcaggacag gagggtggaa gccaaggagc 9960

ccagaggcag aggcagggac aggggaggca caggggctag gctcagagcc acctgatggc 10020

gctggggcac ctgctggcgg ggagcagggc tgtggtcagc agcggagtgg aggggagagc 10080

tgtgctgagt gcacagatgg gaggagggaa gagtccaggg aggcccagaa aggcccagag 10140

tgcagcaggc ctggggcgag gggaggggtg aggctccgtg cgttcaggga gctgacccag 10200

cagagcagag gccactgagg agctgaggtt ctggagaggc ttccagagca ggagcagtgc 10260

agggacggga ggatctggga gctcacccag gaggggcaca tgggcaaggg caaggggctc 10320

tgttggggag acctgactgg acactggggc tgctccacag catagggaac aagccaagtg 10380

ctgcaaaaac aaaaatgagg ccagaaaaac agcccaaacc tggacagagg gtgccaggac 10440

aggcaggggg gcaacagtga cctgagtgac attgctgccc gggttgaggg agggcagagt 10500

gagcaggggg caggcattgg agttcagggt accaggaccg agcagccaca ggtgagcagg 10560

gcaggtgggg gcagaaggag cagggggcac ctcctggagc tcagcagacc agggcagagc 10620

aactgaaggt gaacaagggc aggtgggagg caggatgagc agggggaaga ccctggagct 10680

caggggacca gggcagagca gcctcaggtg cctcaggtga gcaggggctg gtgggtggca 10740

ggacgagtag gggacagctc ctggagctca ggggaccagg acagagcatc aagagctgag 10800

catggctagt gggaggtggg cgaacagggt gcagcccctg gaactcggga ccagggcaga 10860

gcagcggcag gtgagcacgg gctggtggga ggcaggagga acagggggca gctctgggac 10920

ttcaggggac caggggaggg catctgaagg tgaacagggg ctggtggggg caggaagagc 10980

agggggaagc ccctggagct caggggacca gggcagagca gccacaggtg agcaggggct 11040

ggtaggaagc aggaggagca ggggacagcc cctggagctc agagcaccag ggcagagcac 11100

cctcaggtaa gcaggggcag gtaggaggca ggacgagcag gggacagccc ctggagctca 11160

ggggacagag gagagcatca gaaggtgagc aggactgagg cttagcctca gggaatcaga 11220

gcagagcagc cacaggtgag cagggccggt gggaggcagg acgagcaggg gacaggcact 11280

agagctcagg gcaaggcaac cacaggtgag cagggccggt gggaggcatc actcagctcc 11340

tagattttgg caggagctgg gtagttgctg gcagcagaca gctgagggct ggtgaaagtg 11400

cagtgcagcc tcctggtgcc aggaagggag tgtgagccca tcccactgag cagttggcaa 11460

gggtgagctg ggatggagaa gggaaggcat tccagggctc ggggctgagc tctcaggcag 11520

gggcaggtgt ggctgcaggg ggaatgtgtg cttgagacca ggagggtccc agggctggcc 11580

ccagcggacc ctaggcagga aggcctctga ggctggcgcc ccagaaggag caagatgggc 11640

tgccaggagc cagggccacc agcacaatga agctgagtgg aggtggtgca gggcagtgta 11700

gcagcagagg gctgccagag gggcccattc agggcctggg cagagtcagc cagagcctgt 11760

ggtgcaggtg aggggaaggg gtggtgagcg gggccctggg gctgagcaga ggggatggcc 11820

tggctgaggg cagggcactt agcctcctca gaggtcaggg gcacacccca cctgcagtgg 11880

gactccaggg ccactgggcc agcggcagag agaaatgggg cctccctgtg gcctgggggt 11940

cctggcacca cgcagggtgg ggagggccaa gggcaggtgc aaggctccta cctgtgctgg 12000

ggggcctggg ttgagcccag cagggacctt gccgggggaa gctctggaga gagggaggag 12060

gtgggctggt ggccgagaag gccaggccag ggctgggagg gtgaggttgt ggtgactgag 12120

cctccagaag taatgcagga cactgggagg cagggggcat ccaggcactc agggccctga 12180

cctgggctgc tgcacactgg ggctaagggg aaaggagggg agaggctgag gaggaggctc 12240

caggaggcta ttccaaggca gggggttccg gggccctggg gctgaagggc gccgacccta 12300

tgcagtgtct ggcccctctg ctgcacagaa gaaaagggcc ttggagggca gagggcaggc 12360

tatgaccagg gccctgggca agtcaggccc actcactagc ggagggccac gctggggcgg 12420

cagggtcagg agcttcaggg gactcggggg acccacgaga agccatctga gaacagtgtc 12480

cactggtcaa gccaggcacc cataaaaggc tggagtgggg ccaatgggca tgagccgtcc 12540

ctgaggtggc accgatggcc agagctgagg ccaagctaga ggccctggac tgtgctgact 12600

cccggcaggc acagagcgct gacctggctg ccgagccccg cctcctaggc tgcaggggtg 12660

cctgcagaag ggcaccacag ggccaccggt cctgcaagct ttctggggca ggccgggcct 12720

gactttggct ttggggcagg gagggggcta aggtgacgca ggtggcgcca gccaggtgca 12780

cacccaatgc ccgtgagccc agacactgga ccctgcctgg accctcgcag atagacaaga 12840

accgaggggc ctctgcgccc tgggcccagc tctgtcccac accgcggtca catggcacca 12900

cctctcttgc agcctccacc aagggcccat cggtcttccc cctggcgccc tgctccagga 12960

gcacctccga gagcacagcg gccctgggct gcctggtcaa ggactacttc cccgaaccgg 13020

tgacggtgtc gtggaactca ggcgctctga ccagcggcgt gcacaccttc ccggctgtcc 13080

tacagtcctc aggactctac tccctcagca gcgtggtgac cgtgccctcc agcaacttcg 13140

gcacccagac ctacacctgc aacgtagatc acaagcccag caacaccaag gtggacaaga 13200

cagttggtga gaggccagct cagggaggga gggtgtctgc tggaagccag gctcagccct 13260

cctgcctgga cgcaccccgg ctgtgcagcc ccagcccagg gcagcaaggc aggccccatc 13320

tgtctcctca cccggaggcc tctgcccgcc ccactcatgc tcagggagag ggtcttctgg 13380

ctttttccac caggctccag gcaggcacag gctgggtgcc cctaccccag gcccttcaca 13440

cacaggggca ggtgcttggc tcagacctgc caaaagccat atccgggagg accctgcccc 13500

tgacctaagc cgaccccaaa ggccaaactg tccactccct cagctcggac accttctctc 13560

ctcccagatc cgagtaactc ccaatcttct ctctgcagag cgcaaatgtt gtgtcgagtg 13620

cccaccgtgc ccaggtaagc cagcccaggc ctcgccctcc agctcaaggc gggacaggtg 13680

ccctagagta gcctgcatcc agggacagac cccagctggg tgctgacacg tccacctcca 13740

tctcttcctc agcaccacct gtggcaggac cgtcagtctt cctcttcccc ccaaaaccca 13800

aggacaccct catgatctcc cggacccctg aggtcacgtg cgtggtggtg gacgtgagcc 13860

acgaagaccc cgaggtccag ttcaactggt acgtggacgg cgtggaggtg cataatgcca 13920

agacaaagcc acgggaggag cagttcaaca gcacgttccg tgtggtcagc gtcctcaccg 13980

tcgtgcacca ggactggctg aacggcaagg agtacaagtg caaggtctcc aacaaaggcc 14040

tcccagcccc catcgagaaa accatctcca aaaccaaagg tgggacccgc ggggtatgag 14100

ggccacatgg acagaggccg gctcggccca ccctctgccc tgggagtgac cgctgtgcca 14160

acctctgtcc ctacagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 14220

gaggagatga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctaccccagc 14280

gacatctccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacacct 14340

cccatgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 14400

aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 14460

tacacacaga agagcctctc cctgtctccg ggtaaatgag tgccacggcc ggcaagcccc 14520

cgctccccag gctctcgggg tcgcgcgagg atgcttggca cgtaccccgt ctacatactt 14580

cccgggcacc cagcatggaa ataaagcacc cagcgctgcc ctgggcccct gcgagactgt 14640

gatggttctt tccgtgggtc aggccgagtc tgaggcctga gtggcatgag ggaggcagag 14700

cgggttccac tgtccccaca ctggcccagg ctgtgcaggt gtgcctgggc cgcctagggt 14760

ggggctcagc caggggctgc cctcggcagg gtgggggatt tgccagcgtg gccctccctc 14820

cagcagcagc tgccctgggc tgggccacgg gaagccctag gagcccctgg ggacagacac 14880

acagcccctg cctctgtagg agactgtcct gtcctgtgag cgccctgtcc tccgacctcc 14940

atgcccactc gggggcatgc ctagtccatg tgcgtaggga caggccctcc ctcacccatc 15000

tacccccacg gcactaaccc ctggctgccc tgcccagcct cgcacccgca tggggacaca 15060

accgactccg gggacatgca ctctcgggcc ctgtggaggg actggtccag atgcccacac 15120

acacactcag cccagacccg ttcaacaaac cccgcgctga ggttggccgg ccacacggcc 15180

accacacaca cacgtgcacg cctcacacac ggagcctcac ccgggcgaac cgcacagcac 15240

ccagaccaga gcaaggtcct cgcacacgtg aacactcctc agacacaggc ccccacgagc 15300

cccacgcggc acctcaaggc ccacgagccg ctcggcagct tctccacatg ctgacctgct 15360

cagacaaacc cagccctcct ctcacaaggt gcccctgcag ccgccacaca cacacaggcc 15420

cccacacaca ggggaacaca cgccacgtcg cgtccctggc actggcccac ttcccaatgc 15480

cgcccttccc tgcagctgag gtcacatgag gtgtgggctt caccatcctc ctgccctctg 15540

ggcctcaggg agggacacag gagatgggga gcgggtcctg ctgagggcca ggtcgctatc 15600

tagggctggg tgtctggctg agtcccgggg ccaaagctgg tgcccagggc aggcagctgt 15660

ggggagctga cctcaggaca ctgttggccc atcccggccg ggccctacat cctgggtcct 15720

gccacagagg gaatcacccc cagaggcccg agcccagcag gacacagtat tgaccaccca 15780

cttcctgtcc agagctgcaa ctggaggaga gctgtgcgga ggcgcaggac ggggagctgg 15840

acgggctgtg gaccaccatc accatcttca tcacactctt cctgctaagc gtgtgctaca 15900

gtgccaccat caccttcttc aaggttggcc gcacgttgtc cccagctgtc cttgacattg 15960

tcccccatgc tgtcacacac tgtccccatg ctgtccccac atgtccctga cactgtcccc 16020

catgctgtcc ccacctgtcc cggacactct cctccgcgct gtcttgacct gtgcccaaca 16080

ctgtccccca cgctatcccc ccatccccaa caatgtcccc cacagtttcc tcctgtcccc 16140

tatccccgac actgtcctcc acactgtccc cacctctccc tgtcactgtc gcccatgctg 16200

cccccacctg tcccaacact ttcctccaag ctgtcctcac ctgtccccaa cactctcccc 16260

cacactctct ccacctgtcc ctgacactct cccccatgct gtccccacct gtccctgatg 16320

ctgtcctcca cactgtcccc acctctccct gtcactgtcc ccatgctgtc ccctgtccct 16380

cacactttcc tccatgctgt cctcacctgt ccccaacact ctcccccact gtctccacct 16440

gtccctgaca ctgtccccca cactgtcccc acctgtccct gatgctgtcg tctgtgctgt 16500

ccacatactg ttggtgacct ggctctgttc tccaagttca agcctcagag caggcagtgg 16560

tgaggccgtg gcacctgggt ggcctgaggg gtgggcgggc cttgggggca gggctgtggc 16620

ctcgctcacc cctgtgctgt gccttgccta caggtgaagt ggatcttctc ctcagtggtg 16680

gacctgaagc agaccatcgt ccccgactac aggaacatga tcaggcaggg ggcctagggc 16740

caccctctgt ggggtgtcca gggccgccca gaccccacac aggagccgtg ggccatgctc 16800

agccatcacc caggccacac ctgcccccga cctcaccgcc ctcaacccca tggctctctg 16860

gcctcgcagt cgccctctga ccctgacacg ccccccttcc agaccctgtg catagcaggt 16920

ctaccccaga cctccgctgc ttggtgcatg cagggcgctg ggggccaagt gtcccctcag 16980

caggacgtcc ctgccctccg gcccgccagg tgctcacaca aaaggaggta gtgaccagca 17040

tcccaggccc ccactcaggc aggacctcgc cctggagcca accctgtcca cgccagcctc 17100

ctgaacacag gcgtggtttc cagatggtga gtgggagcat cagtcgccaa ggtagggaag 17160

tcacagcacc atcaggccct gttggggagg cttccgagag ctgcgaaggc tcactcagac 17220

ggccttcctc ccagcccgca gccagccagc ctccattcca ggcactcccg tgaactcctg 17280

acatgaggaa tgaggttgtt ctgatttcaa gcaaagaacg ctgctctctg gctcctggga 17340

acagtctcag tgccagcacc accccttggc tgcctgccca cactgctgga ttctcgggtg 17400

gaactcgacc cgcagggaca gccagcccca gagtccgcac tggggagaga aggggccagg 17460

cccaggacac tgccacctac cacccactcc agtccaccga gatcactcgg agaagagcct 17520

gggccatgtg gccgctgcag gagccccacg gtgcaagggt gaggatagcc caaggaaggg 17580

ctgggcatct gcccagacag gcctcccaga gaaggctggt gaccaggtcc caggcgggca 17640

agactcagcc ttggtggggc ctgaggacag aggaggccca ggagcatcgg ggagagaggt 17700

ggagggacac cgggagagcc aggagcgtgg acacagccag aactcatcac agaggctggc 17760

gtccagcccc gggtcacgtg cagcaggaac aagcagccac tctgggggca ccaggtggag 17820

aggcaagacg acaaagaggg tgcccgtgtt cttgtgaaag cggggctgct ggccacgagt 17880

gctggacaga ggcccccacg ctctgctgcc cccatcacgc cgttccgtga ctgtcacgca 17940

gaatccgcag acagggagac tcgagcggga gtgcggccag cgcctgcctc agctgtcagg 18000

gaggactccc gggctcactc gaaggaggtg ccaccatttc agctttggta gcttttcttc 18060

ttcttttaaa ttttctaaag ctcattaatt gtctttgatg tttcttttgt gatgacaata 18120

aaatatcctt tttaagtctt gtacttcgtg atgggagccg ccttcctgtg tccacgcgcc 18180

tcctgccccc ggtgggaagc acggtcagga ggaggctggt ccagctgcac ctcgggggct 18240

ccctgcatac gccccccgcc tcctgcagcc acacgcattg cccgagcgac cctccctggc 18300

ccctgtcgct acatggaccc ccggggtttc tcctcttttc tacatggatg cagtttctcc 18360

tcctgctggg cacggtgctg cctgccctgg tcactctgcg ggggacaggg cctccaggga 18420

aagctgggtc gaggctggga gctggctcag gctgcccagg cagagccaca gggagggcct 18480

tccagaacca accatggtcc gaagcgagag gtgggtgtca gatctgtgtg agtcagctca 18540

ggaccacagc ggggcggctc ccacagcaga catggatcct cccaggccta gagaccagga 18600

atctgagatc aggatgcagg cagggctggt ttctctcaag ccctctctcc ttggcttgta 18660

gacaccgtct cctccctggt cctcacatgg ccatccctct gtgtgcccgt gtcctaagct 18720

tctcttctta taagaacaca catcggatta gattagtgac cccctatgaa cttaatgacc 18780

tctgtaaaga ccccatctcc aaatagtcac attgtgaggc cagggattaa gacttgaata 18840

tatgaatttg taggggccac gatttaaccc atcacagtcc agactctggc ccccaaaatt 18900

catgttcttc tcacatgcaa aacacattca tcctgtctca gcatccccct gggcactagg 18960

tcatgtagca aggacggatt ttcaacagaa ataactattg caacagaaga aagagtccgg 19020

catgacctgg actcaccttc atctgtgcag aggccacagc cttgtaaagg gaggtggtag 19080

ggggagcagg gagggtgctc ggggctcagt cgtcggggaa gggaaaagtt gcccagcgct 19140

ggtcagcgtc cccgggatgg gacccgctgt gtctgtgccg gccactgttg aggtcaggat 19200

tctgtcctcc cagagcctgg agacacaggc cccatccttc ccaatgggga cacttcaggg 19260

agtggctctc aggtcccgag aaagaccctc ctgggtcaca ggaaatgcac agacatcggg 19320

aacggataga aggtcgtgtg gttgcggccc tctcagcaga taccctgaga aagggaggtc 19380

ggggttggtc caaacggtga gttctggtgc acggagcttt ctcaggcagg tgttgacggg 19440

gcaggggtcg gcctaggggt acggccagaa gctgttagaa actgttagtg tctgctcaag 19500

tctttacaag ccaaggttga ggccgagtgg agaggctccg aggagcctgg ctggaactca 19560

gtcaaggaca gggtcttgtt actgcagtgg ctgcggtggc tgcggtggct gcgaaatgcc 19620

gtcggagttg cctgtggcag gagagagacc atctcaccca ggaaggagga gtggttggat 19680

tcgtttgtgt ggcatcgagc agctggagct tcaccaaaca cagagttggg gactaaatcc 19740

ccagactcca ggccctgcca tgccgtggga aggctcgcca ctggagggtg cgctccaggg 19800

ggcctggcct gaactgggtg ctgaagccca gccctttaac tctcaggaca cgctgctgca 19860

gccccgcggg gggtgaggga gagagcacct ggggtgcagg gcgggcagct gctgcatcac 19920

cggctctatc ccaagcccaa ggatggcgtc ccagagatgc aggagagctt tgtccagaga 19980

aggtgccagc cctcagggac cctgctggag agatctccac cctctgccct tcaaggggcc 20040

ctacgggcct ccaggtgctc tggtggggtg ggctccagtc cactgtctga ggatggacgg 20100

cctggccagg ataaggaaag gaaacccagg acggtgccgg gctccgggtc attccgtgca 20160

ctgagcaggc tgagttggga agaagcagat gcttcctgca gctgctgccc cctgcagggc 20220

ctggcgcctg gaccaggttc ccctggggaa attgggcccc tccctgagcc acccggggcc 20280

cacctcccac tttctacctg ggaccgagca tcctccagag ggtcagccct cctgcgggaa 20340

caccatgccc agccccagga ccctccctca actctccagc aaggctgccc ctgcacaccc 20400

cccagcagcc catgctgtga tgtaacatga catcgtgtga catggtgtga tgtcctatca 20460

cagtgtgaca tccctggtgt gatgtggtgt gacatggtgg gatgtggtgt gacatggtgt 20520

gatgtccctg gtgtgaggtg gtgttggaca tagtgtgatg tgatgcgaca gatgtaacat 20580

ccctggtgtg atttggtgtg acgtagtgtg atgggtatga catccctggc ttgatgaggt 20640

ataatatgaa atgatgtgga atggtgtgac atgatgtgat gtgatgggac agggtgtgac 20700

gtccctggtg tgatgtgatg taacaggctg tgacatagtg tgatgtgggt ggtgtgacat 20760

gatgtgatgt ggtgtgatgg tgtgggacat ccctggtgtg atggggtgtg attaaacctt 20820

gattccatgc agcacatgtt tctgtgagca cagggttggg gctaaagtta caggttaaca 20880

gcatctcaaa gcagaacaat ttttctttgt acagatcaaa atggagtttc ttatgtgttc 20940

cttttctaca taggcacagt aacagtctga tctttctttt ccccacagtg tgacatggtg 21000

tgacatttct ggtgtgactc ttgtgttgtg acatttgtgg tcaccccagg atacagaggt 21060

ctctgtggcc aagggaaggg ggagaatgga accatctgag catgttgacc tggaggaatt 21120

ggtggccctt gagtccacga agcccaccct tccaggtgcc cctgccccac gtgacccaag 21180

tgggcttgca gagcagcaag caggactctg gttagacagg aggaaggacc tgccaccacg 21240

tggccttgtg aggagacaga gcgaggctgt gacctcggcg tccgcccagc acagggtgct 21300

gctgaagctc ctccggtcct ctcagcagcg gtctcagagc aaggcccaag gcaggctgaa 21360

gagaggggca gagggaggat gctggggagg caggtgtgag gggactgaga gcccaggttt 21420

cagctgagcc cctccacagg gaaggagcct agctgaacac ccatctcccc acacgctccc 21480

aaccctgcct ctgcccgacc acctcccaga gggcacctcg aaccctctgc tacccacact 21540

cagcaagggg tatggtgccc ccaccaagcc cagccagcaa agcctggcac agccacgcct 21600

gtgcccacca ctcccatggc caaggtcact gctaacatgg caggacagag ccaggcctgg 21660

aggagacaga acatcagtcc catggggaag ctccctgctc acacggcagg gccaggcctg 21720

gagaagacag aacaccccat ctggcatggt actcaggctg cacatgcctg ccacgaacgg 21780

gggccacgcg acaatgcctg ccacacatgg gggccacgca acgatgcctg ccacacatgg 21840

gggccacgcg acgatgcctg ccacacatgg gggccacgcg acgatgcctg ccacgcatgg 21900

gggacacgtg acacacacat acacacacgg gcctcacagg cacacaaatg cttgcgaacc 21960

cagcacccac ccagcacact caggcatagg ctccctgggc aggtcacaac ctcacgcctt 22020

gagctagtcc atgtgccagg cccgtcaccc acgtactcgt ccccggtgct agctattgag 22080

ccacaccgtc ttctctgtgg atccctccca gcccactcag cacaatggac atgctctctc 22140

ccgtccagtg actgcaccgg cctcttcccc tcagcaccca agtctggcca tctcccagga 22200

aacccggacc accacgggca gggaccacgt tcctcactgt ccacgtggac caccccacac 22260

ctgaccccag atgcagtcac actacgtcct gcttcaatat tgaaaagggg aaaagctgga 22320

ggagggtaaa gatgaaagag aaaaaagcaa gaggggaggg tcacattctt ctgaggcttt 22380

gattacatct cactgagccc ccacgttgca tgaaaaggag gggtggaggg agcaattaca 22440

catttgcctt gtgctcagta aatctgcact ttataagcaa ataaacagag tagaggaaga 22500

agtcaaatat gcattcgtct caggggcagg agggatgatt tcttgtctca ttttgtccca 22560

tgtcatgaag accgggctgt taatttatat tgtcagggtg agggaggcca cctgggtaga 22620

cctggcctat ctgctgctgc tctcagtttg gaaacaaaag gaaacgcatg actttttttt 22680

tttttttcat gactcagctt cccacctcaa ctgttctttt tggcatagtg actttagggt 22740

cctgagattt tattttcctt tcacaatggt caagatcaca ccctcagtat tcaggagagc 22800

tggtccaacc cagacccctg ccgtccctgg acgcttttca aatacgtgct gcagtctctg 22860

acactctctc accatggtag aaactgaggt cttggtgagt ctgcagcccc tgctcatggg 22920

gacaatgcag gctgaaaccc gcatttctga ccccaaagct cctgttcctt tcactcaccc 22980

ccacaccagc tctttgagtc cagagctttg tccttgcctg agtcctaccc tcagggacag 23040

gggcccaacc cagccaccaa cacatcatac cctgagaggg tgccaggagc ccagagatgt 23100

ttggagagca cagtagccct ggaagctctg tggagatgct gcacatttct ctattcaaca 23160

gatactcgcc aggtggccag cagcagagat gccacatggc atggagctcg ccttggccaa 23220

caggacaggt gtggggttgg gaggccttcc cagggcactc cttgaagcag agctgtgggg 23280

aaaagttggg gccaatctca gatctccccg tcaacaccgg gtctcctgcc ctcctgggcc 23340

acagaaaact aagctccgtg gatactgcgg ctgggtgggg ccgtggggga gaagaaatca 23400

caacgagtta aaagatcatt ttctaaaact gttatgatca ggactcacat aaccatatga 23460

cgacacattt cagagatgct ctttatctca ttaattaagg tgtcgtaacc agttcaaagt 23520

ggaattctaa gtactacact tacataattg attcaggaat gctaaaagga gttcatagat 23580

agatgcaaaa ctggcctttt ccctggaaga tgaggagcaa ttcattgtcc ttccaaagat 23640

gagaacttga atttctacca actcaaagag cttttgcatt gctatcaatt atgtacaact 23700

tagagcagtg gtccccaaca tttttggcac caggaaccag ttccatggaa gacaattttt 23760

ccacagacca ggatcggggg atggcttggg gacaaagctg ttccacctca gatcattagg 23820

cattagggtg tcataaggag tgtgcaactt agatcccggg aatgtgcggc tcgcaatagg 23880

gttcgctcct gtgagaatct aatgctgcca ctgatctgac aggaggtgga gctcgggcag 23940

gaatgctcac acacccctca cctcctgctc tgtggcccag ttcctaacag gccatgaacc 24000

ggttccagtg catgacccag ggattgggga cccctggctt atagaggtgt aaaatagttc 24060

aaaggaaata aaagatgcag agctccacag aatgaaataa cctggaagag tgtacaagac 24120

gatgccttgc tttccatgga aggcacctac taatttttct caatgtttcc tataaacata 24180

tataactgac tgacagaaac agatccataa tataaagaag acccctgtaa accaatgaga 24240

aaaaaaatca aataatccaa cgaggaatag gcaagagaat tgaacagatg ttttacagaa 24300

gatatccaaa tagccactaa acatatgaaa aggtgttgaa ccacactagt caacagggaa 24360

atgaaaatga aaaaccacat gagagaaagt agttctgatt ccagtaatgc tagagcagct 24420

aatatcagac tagccctttg gcagatggca attataaaca ctggaaacac tgtaagcaca 24480

cacaacaccc acacacacca attgcaggca ctggaacatg accagaagta ggcaaacact 24540

agtaaggatt attccgtgaa atattcgtct gaagtcacac cccagtgcat gtaatgggtg 24600

cagctagagt ttaagcagga aactgcagcc ctcctggtga ggagtgggat gcagggctgc 24660

attttcagag cagctggaaa tgaagagaag atgtccataa aggagaagct caccgaaggg 24720

aaaccacaca atctgcaagt aaactccact gaaacctctg gccgatccct taggtgtgca 24780

tgggtaggga aaactccaaa gggcccagca gaaagcaaca cctgtaagtc gagagaactg 24840

agattccagc tactgccaac tgccaggcag acagacttgg gagtttgagt caactcaagc 24900

taactgctaa cattgaaaaa aacaattaat gctctgctaa gaaagaatgc aaaccccata 24960

gcctgtacgc atgttatcaa catcaggtgc acatccaaaa ttaccatgga tgcaaagaaa 25020

catgaaaatg tgatccatag tcagaagaaa aagggatcaa tggagatcaa ctccaagatg 25080

acctagatgc agatacagtt atcagacaag gactttaaag aagttatgtt aaacatgttc 25140

aaagacttaa aggaaaatac tattataatg agtgactaga tggggaatct ctataagaga 25200

cggaaaatat ttacaagaac caaatgaaaa ttctagaact gaaagtatga ttctgaaatg 25260

aaaaacatca tttttcagag cagggtgaga atggacatgg gactcagagc tgagcaggcc 25320

tggtgggccc caggagggag acacagagga ctgggggatt tcaaggctgg cagaggccag 25380

agatggatcc ccagctggga ctggacctgg gcttatggga gcaacaggtg acccatcctc 25440

cttcctgggg gcccaccctg cccggcccct ccagcccagc acaggcattg gatagaaccg 25500

ggagagagca ggccaggcac tgaggcctct gccccaaatg cccacagcct ggggaaaatg 25560

agcagataga tgggggggca agtggatccc caggcacacc cacacagtgc acacagcccc 25620

acctgggcca gagggggcag gaggctcgcc acccctgctg tggtttctcc cacacttgat 25680

gcaggtgata ttcctctgag attgtggact aagagttggt gctggaaggg gttagccatc 25740

ttggagatgt tgctatgggg tgcagggatt ttgcatgtga gaaggacatg attatggggg 25800

gagcggaggg caaactgttg tgggttaaaa tgtgtcccct ataaattcat gtgttgaagt 25860

cctaaccccc aggaccacag aatgtgacct tgtttggaaa cagtctttgc aactgcaatc 25920

aagttcagat gaggtcaccc tggagtaggg caagcctctg atccaatatg actgctgtcc 25980

tcatgaaaag ggggaatctg ggtacagaca gcacgtgggg agaacaccct gtgaagatgg 26040

tgctgcttcc ataagccaag agcagcagag acggccggca aagcccagca gcaaggagag 26100

agcctgggac agagtctccc atgacacaga ggtgccagcc ccgccgaggc ctccatccca 26160

gatgcccggc ctccagaacc aggacggaat aaacgtctgt tgtttaagcc acgcagtctg 26220

gggtgctgtg ttgccagggc cacagttaac ggatacgagt gttgtcctga gctgccagcc 26280

ccacaggctg cacgaggcct ccctgcccca gcccagtgca gactccccag ccccctgggt 26340

gtgccatggg cagtgcgggg cccctcactg catcctcccc cagcctggga ggttgagccc 26400

attatgagct ccatggggtg aagccggagc cagaagctgg gagccgactg ggagcctgcg 26460

gctggaggat ggatttcccc agggacccac acgtgcacct ccacctgtct cctggacatt 26520

ctctctgagg gcagggctgg tgtcagctca gggatccagc agggacacaa gggtgggccg 26580

ggtccttgtg gagagcacat ttagtgggag ggacatgatt tcccttcaaa gtgcccattc 26640

tggatgcttc ctggtccacg ctggacactt cctgttccac gctggacgct tcctgttcca 26700

cgctggacgc ttcctgttcc acgcttgatg tttcctgttc catgctggat gcttcctgtt 26760

ccatgctgga catttcctgt tccactctgg atgctccctg ttccattctg gatgcttcct 26820

gttccatgct ggacatttcc tgttccactc tgcatgcttc ctgttccact ctggatgctt 26880

cctgtgcgaa acctcctcgg gcttttggtc tgcccagtcc ctctggctgc atctcgtccc 26940

ccgctacctc ccacctccac atccgtcctt gcccagctcc tctctctctc cagagtttcc 27000

acctggcaag gtccctgatg agctcagtcc aggctccccc agcacaggta ggagcctagc 27060

acctgccctt ggacctcccc accctgcatg atgccagcat ccccaggccc cagggaggcc 27120

ccatttctct ctctactgct ggcccagtgg ccctggagtc ccactgcaac tcgggtgtgc 27180

ccctgacctc tgaggaagtt aagtgtcctg tccctagcca ggctatcccc tctgctcagc 27240

cccagggccc tgccccttac cccttcccct cacctgcacg ataggctctg gccaactctg 27300

cccaggccct gaatgggccc ctctggctcc cctctgctgc tacactgccc tgcaccacct 27360

ccactcagct tcagtgtgtt catccacctg tcccaagtcc cctcggcccc caggagcaca 27420

gctggtggcc ctggttcctg gcagcccatc ttgttccttc tggagcacca gcctcagagg 27480

ccttcctgtg cagggtccac tcggccagcc ctgggaccct cctggtctca agcacacgtt 27540

ctccctgcag ccagacctgc ccctgcctgt gagctcagac ctgagccttg gaacgtcttc 27600

ccttctccat cccagctcgc ctttgccagc tgctcagtgg gatgaactca cactcccctc 27660

cctccaccat gagtgagagt cagctggaga gatgcccagg ccaaagcagc caccagggcc 27720

cagtgggggg ccagaagctt caggtgagag gcccaggtat tgagaggctg agaccatggg 27780

cagaatggtc ataatcgctg ccagtctcag tccagcccca gggactcaga gacagagaaa 27840

agagcagcac acaaggtccg ggctccccac cttctcccgt gagtatgggg gagtatgggg 27900

gcagccacca cccccatccc cacacaccca tgaggcagcc tcggctctgt gtggactccc 27960

cctcgccctc tgacacagaa accaccagaa gaaaagggaa cttcaggaag taaggggtgc 28020

cgctggtttc aatcctgttc ttagtctttg cagcgtggag ttcacacccc tggggacctg 28080

ggacctgagc tgtgatttcc taggaagaca aatagcggct gacggcgggg gcggggccgc 28140

ccacatgtac ctcgccagaa caggaagggt tgagaccccc acctcggtga gtggggtcag 28200

cacagggcag gggcacaggc tcgggaggag gacagcctgg gcgcagccgt cggcgctcct 28260

agacctgagc tgctgaacag gctgcaagag gctggggaga cgcgggcgcg aggccagccc 28320

cacatggaag cccaagcgga gccagcacgg gggaggtggg cagccttcag gcactgatgc 28380

ccacccagtg cgagacgacg gggaccgtgg gcaggggctt ccaagccaac agggcaggac 28440

acaccagagg ctgactgagg cctccaggac gaccgggctg ggagcacgag gaacatgacg 28500

ggatgcggca gaaccggctg tggggtgatg ccaggatggg cacgaccgac ctgagctcag 28560

gaggcagcag agcgagggag gaggagaggc cccaggtgaa cggaggggct tgtccaggcc 28620

ggcagcatca ccagagccca gggcagggtc agcagagctg gccgtagggc cctcctctca 28680

gccaggacca aggacagcag gtgagccggg agcagagcag cgagggtgag tgtggcagca 28740

ggacagaagg gtggaagcca aggagcccag aggcagaggc agggacaggg gagggacagg 28800

ggctgggctc agagccagct gatggggctg gggcacctgc tggcggggag cagggctgtg 28860

gtcagcagcg gagaggaggg gagagctgtg ctgagtgcac gggcgggagg agggaagagt 28920

ccagggaggc ccagaaaggc ccagagtgca gcaggcctgg ggcgagggga ggggctgagg 28980

cccagcagag cagaggccac tgaggagctg aggttccgga gaggcttcca gagcaggagc 29040

agtgcaggga cgggaggatc cgggagctca tccaggaggg gcacataggc aaggggctct 29100

gttggggaga cctgactgga cactggggct gctccacagc atagggaaca agccaagtgc 29160

tgcaaaaaca aaaatgaggc cagaaaaaca gcccaaacct ggacagaggg tgccaggaca 29220

ggcagggggg caacagtggc ctgagtgaca ttgctgcccc gggttgaggg aggacagagt 29280

gagcaggggg caggcattgg agttcagggt accaggaccg agcagccaca ggtgagcagg 29340

gcaggtgggg gtagaaggag cagggggcag ctcctggaac tcaggggacc agggcagagc 29400

agccacaggc aaacaggaga ggggaggggg ggcaggagga gcagggggca gctcttggag 29460

ctcaggggac cagggcagag acgccgcagg tgagcagggg caggtggggg ggcaggagga 29520

gcagggggca gctcttggag ctcaggggac cagggcagag cagccacagg tgagcagggg 29580

caggtggggg gcagaaggag tagggggcag ctcttggagc tcagaggacc agggcagagc 29640

agccacaggg gaggaggggc aggtgggagg caggatgaac agggggcggc tcctggaact 29700

caggaaacag gggagagcat cagaaggtga gcagggccag tgggaggttg cagagcaggg 29760

gacagctcct ggagctcagg ggaccagggc agagccgccg caggtgagca ggggcaggtg 29820

gggggcagga ggagcagggg gcacctcctg gagctcaggc gaccggggca gagcagcctc 29880

aggtgaaaag ggccggtggg gggcaggagg agcaaggggc agctcctgga ggtcagggga 29940

ccagggcaga gccgccgcag gtcagcaggg ccggtgggag gcaggacgag caggggacag 30000

gcactagagc tcagggcaag gcagccacag gtgagcaggg ctggtgggag gcatcactca 30060

gctcctagac tttggcagga gctgggtagt tgccggcagc agacagctga gagctggtga 30120

aagtgcagtg cagcctcctg gtgccgggaa gggagtgtga gtccatccca ctgagcagtt 30180

ggcaagggcg agctgggatg gagaagggaa ggcattccag ggctcagggc tgggctctca 30240

ggcaggggca ggtgtggctg cagggggaac gtgtgcttga gaccaggagg gtcccagggc 30300

tggccccagc ggaccctggg caggaaggcc tctgaggctg gcgccccaga aggagcaaga 30360

tgggctgcca ggagccagga ccatcagcac aatgaagctg agtggaggtg gtgcagggca 30420

gtgtagcagc agagggctgc cagaggggcc cattcagggc ctgggcagag tcagccagag 30480

cctgtggtgc aggtgagggg aaggggtggt gagcggggcc ctggggccga gcagagggga 30540

tggcctggct gagggcaggg cgcttagcct cctcagaggt caggggcaca ccccacctgc 30600

agtgggactc cagggccact gggccagcgg cagagagaaa tggggcctcc ctgtggcctg 30660

ggggtcctgg caccatgcag ggtggggagg gccaagggca ggtgcaaggc tcctacctgt 30720

gctggggggc ctgggttgag cccagcaggg accttgccgg gggaagctct ggagagaggg 30780

aggaggtggg ctggtggccg agaaggccag gccagggctg ggagggtgag gttgtggtga 30840

ctgagcctcc agaagtaatg caggacactg ggaggcaggg ggcatccagg cactcagggc 30900

cctgacctgg gctgctgcac actggggcta aggggaaagg aggggagagg ctgaggagga 30960

ggctccagga ggctattcca aggcaggggg ttccggggcc ctggggctga agggcgccga 31020

ccctatgcag tgtctggccc ctctgctgca cagaagaaaa gggccttgga gggcagaggg 31080

caggctatga ccagggccct gggcaagtca ggcccactca ctagcggagg gccacgctgg 31140

ggcggcaggg tcaggagctt caggggactc gggggaccca cgagaagcca tctgagaaca 31200

gtgtccactg gtcaagccag gcacccataa aaggctggag tggggccaat gggcatgagc 31260

cgtccctgag gtggcaccga tggccagagc tgaggccaag ctagagacac tggactgtgc 31320

tgactcccgg caggcacaga gcgctgacct ggctgccgag ccccgccccc taggctgcag 31380

gggtgcctgc agaagggcac cacagggcca ccggtcctgc aagctttctg gggcaggccg 31440

ggcctgactt tggctggggg cagggagggg gctaaggtga cgcaggtggc gccagccagg 31500

cgcacaccca atgcccgtga gcccagacac tggaccctgc atggaccatc gcagatagac 31560

aagaaccgag gggcctctgc gccctgggcc cagctctgtc ccacaccgcg gtcacatggc 31620

accacctctc ttgcagcttc caccaagggc ccatcggtct tccccctggc gccctgctcc 31680

aggagcacct ccgagagcac agccgccctg ggctgcctgg tcaaggacta cttccccgaa 31740

ccggtgacgg tgtcgtggaa ctcaggcgcc ctgaccagcg gcgtgcacac cttcccggct 31800

gtcctacagt cctcaggact ctactccctc agcagcgtgg tgaccgtgcc ctccagcagc 31860

ttgggcacga agacctacac ctgcaacgta gatcacaagc ccagcaacac caaggtggac 31920

aagagagttg gtgagaggcc agcacaggga gggagggtgt ctgctggaag ccaggctcag 31980

ccctcctgcc tggacgcacc ccggctgtgc agccccagcc cagggcagca aggcaggccc 32040

catctgtctc ctcacccgga ggcctctgac caccccactc atgctcaggg agagggtctt 32100

ctggattttt ccaccaggct ccgggcagcc acaggctgga tgcccctacc ccaggccctg 32160

cgcatacagg ggcaggtgct gcgctcagac ctgccaagag ccatatccgg gaggaccctg 32220

cccctgacct 32230

<210> 77

<211> 340

<212> PRT

<213> Intelligent

<400> 77

Ala Ser Pro Thr Ser Pro Lys Val Phe Pro Leu Ser Leu Asp Ser Thr

1 5 10 15

Pro Gln Asp Gly Asn Val Val Val Ala Cys Leu Val Gln Gly Phe Phe

20 25 30

Pro Gln Glu Pro Leu Ser Val Thr Trp Ser Glu Ser Gly Gln Asn Val

35 40 45

Thr Ala Arg Asn Phe Pro Pro Ser Gln Asp Ala Ser Gly Asp Leu Tyr

50 55 60

Thr Thr Ser Ser Gln Leu Thr Leu Pro Ala Thr Gln Cys Pro Asp Gly

65 70 75 80

Lys Ser Val Thr Cys His Val Lys His Tyr Thr Asn Ser Ser Gln Asp

85 90 95

Val Thr Val Pro Cys Arg Val Pro Pro Pro Pro Pro Cys Cys His Pro

100 105 110

Arg Leu Ser Leu His Arg Pro Ala Leu Glu Asp Leu Leu Leu Gly Ser

115 120 125

Glu Ala Asn Leu Thr Cys Thr Leu Thr Gly Leu Arg Asp Ala Ser Gly

130 135 140

Ala Thr Phe Thr Trp Thr Pro Ser Ser Gly Lys Ser Ala Val Gln Gly

145 150 155 160

Pro Pro Glu Arg Asp Leu Cys Gly Cys Tyr Ser Val Ser Ser Val Leu

165 170 175

Pro Gly Cys Ala Gln Pro Trp Asn His Gly Glu Thr Phe Thr Cys Thr

180 185 190

Ala Ala His Pro Glu Leu Lys Thr Pro Leu Thr Ala Asn Ile Thr Lys

195 200 205

Ser Gly Asn Thr Phe Arg Pro Glu Val His Leu Leu Pro Pro Pro Ser

210 215 220

Glu Glu Leu Ala Leu Asn Glu Leu Val Thr Leu Thr Cys Leu Ala Arg

225 230 235 240

Gly Phe Ser Pro Lys Asp Val Leu Val Arg Trp Leu Gln Gly Ser Gln

245 250 255

Glu Leu Pro Arg Glu Lys Tyr Leu Thr Trp Ala Ser Arg Gln Glu Pro

260 265 270

Ser Gln Gly Thr Thr Thr Tyr Ala Val Thr Ser Ile Leu Arg Val Ala

275 280 285

Ala Glu Asp Trp Lys Lys Gly Glu Thr Phe Ser Cys Met Val Gly His

290 295 300

Glu Ala Leu Pro Leu Ala Phe Thr Gln Lys Thr Ile Asp Arg Met Ala

305 310 315 320

Gly Lys Pro Thr His Ile Asn Val Ser Val Val Met Ala Glu Ala Asp

325 330 335

Gly Thr Cys Tyr

340

<210> 78

<211> 427

<212> DNA

<213> Intelligent people

<400> 78

acaacaggca ggcaggggca gcaagatggt gttgcagacc caggtcttca tttctctgtt 60

gctctggatc tctggcgcct acggggacat cgtgatgacc cagtctccag actccctggc 120

tgtgtctctg ggcgagaggg ccaccatcaa ctgcaagtcc agccagagta ttttatacag 180

ctccgacaat aagaactact tagcttggta ccagcagaaa ccaggacagc ctcctaagtt 240

gctcatttac tgggcatcta cccgggaatc cggggtccct gaccgattca gtggcagcgg 300

gtctgggaca gatttcactc tcaccatcag cagcctgcag gctgaagatg tggcagttta 360

ttactgtcaa caatattata atttaccgtg gacgttcggc caagggacca aggtggaaat 420

caaacga 427

<210> 79

<211> 121

<212> PRT

<213> Intelligent

<400> 79

Met Val Leu Gln Thr Gln Val Phe Ile Ser Leu Leu Leu Trp Ile Ser

1 5 10 15

Gly Ala Tyr Gly Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala

20 25 30

Val Ser Leu Gly Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser

35 40 45

Val Leu Tyr Ser Ser Asn Asn Lys Asn Tyr Leu Ala Trp Tyr Gln Gln

50 55 60

Lys Pro Gly Gln Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg

65 70 75 80

Glu Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp

85 90 95

Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr

100 105 110

Tyr Cys Gln Gln Tyr Tyr Ser Thr Pro

115 120

<210> 80

<211> 32230

<212> DNA

<213> Intelligent people

<400> 80

gaattcctgt ttctagtagg gacagaaacg gaacccaggc tgttctggcc aatcatgccc 60

tatctcagaa tgttacattt ccagaacatt ctacagttat tccagaaaac tacaatcaag 120

aaagggaagg ggactgggtt gattcaagac aaaatgaaaa ctattctgca aataccatag 180

tgaaatcaca gctcagtaat ctttttgtga ctggcttatt tcatttagcg taatgtcctc 240

tagtttcatc catgttatag catgtgtgag aatttccctt tttaaagcta aataatatgc 300

tattgtatgt atatatcaca tttggattac cagttcactc ctttgtgaac atttgagttg 360

ctctacctgt tggctactat gaataattcg gttctgaatg tgggtataca aatatctcct 420

caagttaatg tcttcaatta cttgggtata tgtccaaaag tggaattgct ggattatata 480

gtatttccat tattaattct tcgaggaatt gccatctggt ttcccacagc aggtgggcca 540

tttacatcac cacgacagtg tccacaggag ttccagttcc ctaagttctc accatgactg 600

gtcattttct gttggaaaaa aaaatcctaa taggtgtgag gtgggttttg tttttatttt 660

tctaaggatt aataatatta agcatctttt cctatgctag ttatctaatt atctctaaag 720

aatattctcc agagaaatgt cgattcatat acttttctcg tttttaatca ggtattttat 780

tttaatgttg acatgtagga cttattttta tgtactagat attattaacc ccttatcaga 840

tatacgattt acaaatattt tcttctattc cacatgttgc attttcactg tgttggttat 900

gtcttttgat gcccatttta catttttatg tagtccaatt tatcttcttt tctacttttg 960

cctgtattat ggtattaaag gtgttagtat tcaaatgtct ataaatactg acttactatg 1020

ctgagaaggt cactctagac catctctctg atggtggagc taagaatttt acactctctc 1080

attttgtacc agggcacagt gcagctaggt gagaacacag tggctttctc gagcttattt 1140

gtcttgcatt tttggcgaca aggattgttg ttcacattgg cctcctctgg caggtccacc 1200

tggaaagcat tatatttagc aagaaaaaag gaggctgtga atgatgtcaa ctgtgtatat 1260

tccctgttgc aagtgtcaaa tccatgaagc ataaaaggat ttactagagg atattaaatt 1320

ccttgcaaag tgttgaaaaa cctgaaggaa taggctccag gcaaagcctc tagaacaatt 1380

ccaagaatgg cactgctggc gcaggctggg gaggagctcc tgctgcctga gactccacat 1440

tcaagctgtc tcctgcagga aagagagcag ctcttctcac tactgccccc tgaaggacag 1500

cagctctgct atcaactact agagacctga cccctttccc tgcagccctg cctgtgttga 1560

ttatatcttc atgtcagact catgtagatt catctgtttt caagggtaca ggggttattt 1620

ctgcccaagt ccacactgtg gctttctatc ataaatacac gtttctacac ttgaatttcc 1680

aggaattact agtatcaaca gcaaaatgct acaacccaac ataaagattt cctttacaaa 1740

gaacatcatg ctccccatgt agcattgtgc tccccaaaat gtggggaaac tccaatctgt 1800

tcatcccaac accatctagg aaaaaatgtt cttctctcat gagccattaa tctgtctact 1860

tattgtttaa tttttggcct aaattgtaaa ggtggctaaa aatagcactt tgaatttttt 1920

taaagaaagg gtatatagaa aatattaaat tatttaaaat acatatatgc atgacacatt 1980

tagttattgt caccagcttc tttcacattt attttattct gtagtttctg gcaatacttt 2040

ctctatggta gagtctggaa atgctactcc tcatgtcctt ggcttcctaa gtcccagatg 2100

tggtgaagaa tgcacatcag ctgtgttact acagaatgct tttggattga gatgggggag 2160

gggacgtgct tgatctcact caaatgtgct caatttcctg gtctgggtca cagccaggca 2220

gcccagtcct atgggcacaa tatcttcact cgccaggttt ctatcaaaat gagagtgtcc 2280

ttcttgactg tggaaaacac ggggcaggga gaggatcaag aaatagcaaa aggagtattt 2340

ggacttgcct gagctgccac tctggtttcc ccaaatgttt tcattccatt tcattcatga 2400

taataaacac ctttctgcct aaccagccat aatcactttt cttagatcca gctgattcag 2460

tgataaaact aaagggaaaa aataaataca gttttcaacg tttaagctga ccaggtggct 2520

gtctctggtg tttatacctc catatcccac tatgcatttc atcttctctt tgcctcactg 2580

gagatccaag tgcagggaat gcttcacgtg actgtgatcc ctagaacttc atcttcatga 2640

ctgttgaaat cttctctgag ttttaccagt ggaaatggca ataaatagaa agattccaga 2700

aggtccccta gggtccatat tttctgtact tcttctctaa aataagtaaa atcaattcca 2760

tttgctcttg ctttatattg gctacatcta tgtgcccagg acacacccta aagccctgtg 2820

ttgacaactt atccagtatt gtattaagtc taaatggtca catgcttatc atatcatttc 2880

ttccaatttc atggaataat tgtcatgtca actctttgga ttaagaacca tcaaccagat 2940

aaccacagcc ctgaataaat agaaacagaa tatttcaagc tgttcagttg gcataacagt 3000

gatgagtcta tagctcattt tcaagatata aatattctat ttatgggagt aaaagcacga 3060

atcatggtta gtgtttcaga gcctccacca catctgacag catagcaatt caacagcaca 3120

aggtttgtgt gccagctgtg cctgactaat aatgtttttt tcccctgtct atagggaaat 3180

ctgcacctgt gcacatgaca agagcatgag ctacactttt ttcacttttt ctgaggtaaa 3240

tacattgctt gatccaaaac cgtattatgg aataatttcc tgactctgat gaaagcattt 3300

ggaaaatcct caaataatat ttgtgcagaa atacaaccaa aaagaaaggc aaattcatat 3360

ataggaaaat aaccagtgct ctcctcatgt tacatgaggt ccactagtat cacctgaccc 3420

cagctgttca tctgatccct gatgtgttgt acaatgttag gggttcagga ggggtcattc 3480

ttcttggcaa atgaggtgtt cagaagagcc agtagccctt taagtctcag ttagttgaaa 3540

ttcataattg agtccccaaa gggacctcca taacattggt tcatgagact ccctggtaaa 3600

gctgggaaag gtgactgact gaagtccatg tgttgaatca tcctcattat taaaagcccc 3660

tgctcattaa tagcattttg attaatattc actgggaaag catttttttt ttattttggc 3720

tcatctttga aaagtctagt catagctttt caatcacttt gtctgcaatc gttttgttgc 3780

atttctttga aggcctgatg atctgtcaaa accaacagcc aatattatta cattgccgtt 3840

atgttgggaa aagctgaatg ttgggagaag ctgaggcagg gcttgcatgt ctgacgtaat 3900

gtaaaagagt cttggaacac gtccggggtc cagggtctaa aaccccttgt ggcctttagt 3960

acaccaagct ctgtgctaaa gggtggaagg ctaccctgac gcaccataat ctaagcccag 4020

ggcataaaat cccttgtgac ttggatagaa tccaggactc gtggctctgg agtgtgtcta 4080

cacttgctgg ctcctttctc tttgctctcc aaggatcgat tgtatcttga gttaaaagaa 4140

cctgctctcc attatctcaa gtagtagagc aaatgctaaa ccatcacagg tgtaaatcat 4200

gtgcttaatg caatgcatcc ttttggcctc cacattctta ccacgtgttt ctttgttgga 4260

ttaccaataa atagcatggg atcccagggc tcagggcctt cttagcctcc atacactacg 4320

atggtccctg gtgcccacat ttctctctca aactgtcttt ttctcaatct ttgactccac 4380

cagactttgt cacccccacg acctggtgtt gggtctgatc accccaaaac tgttatcgtt 4440

gcaatgtgat catcagatat aacccttgca gtttactcat tgttatgatt tttacatttg 4500

cagggcttcc ttgacaatgg cactaatgtg acgccatcca gcatccatta ttgtgagaac 4560

tctatttgga atatcaccat tgtgggcatg aacagaatgg agactttcat aggatgcaat 4620

gtttaatatt caacattagt caaaatttgt tctagtccta cccttggagc tccattgagt 4680

tggaaaaata taaataaatt tattttaaat ataatataaa tgtaaatata aacaaattta 4740

aatataatat aaatataata taaatataca taaatttaaa cataatataa atataaatat 4800

aaataaataa ataaacaaaa aaattttctg tagcaagagt aagaactaca tgtggtccag 4860

aataccatta ttttatggaa ggttacaaaa aattttaaat gtatttaatt cagataagaa 4920

ttttataaat atgcatatat tgcatatatg atagaataaa tgtacttaaa tatgaatact 4980

atatatattt gtagccaaca ttttatcaaa aacatatata ttcacttcta tgtatcttat 5040

atacatgtat gcatactttt ccgtaagata aattttaatt ttttattgaa atactaatta 5100

gttttaattt tgtctttaat tttcttttaa taacttcata atttgattac tggatattca 5160

catttaaatc accaatttaa gaaaaaatac atgtgtgtac atataggtag aggtgtaaat 5220

actgtatcag gaagctttta tgcattactg attggtaact ggttaaatac acaactcagt 5280

gactgataac agtaaacatt tgttttcatg ttcacggatg ctcatgttca caatcctctg 5340

actgatcaag gaagggctca gctgagtgga tcctctgaag gacacagacc tcatcttcag 5400

cctatagata tcaattgtct gaggactaag ctgaacacca gtgactacac atgatacaag 5460

attcttgtgg caggtcacag gagtgaacat cccaaaccaa actggacagt tgaatttaag 5520

tccaataatt tctaacatag cttcagatat ttaaaatata ttcctttatt tcagtgagta 5580

caaattttca agaaaatgtt tactccattt aattatagag gtgtttgatc attccatgga 5640

caaataatta tgttttcatc ctttacaatc ttgaaaatat ttgcaaatgt aaatttgcat 5700

taataagaaa ataaagctgg atgtgttttc aacatgtggc tttaaatata attttttaaa 5760

atggcctcat tgggggaaaa tcattttaac ttatatgaat atcctttttt gcctctcttg 5820

tgttctatag agtggcccaa taagaggtcc tcccatgaga tttggaatca ggaaaggatg 5880

gctcaatatt ctccattggt acctaggaca gacacaggga cagagatgag gactggagaa 5940

acacctggaa agatgctgta ggaagctgag agcatcagca cccccacccc taagcttcca 6000

gacaggactg aggaccacat ggttagatag cccatacttc aggggaagat gcattcagtt 6060

ttctgaggga gcaacagaga ttcctgcttc taatatcaac tttcctgact actatatcct 6120

tggctttgaa aggttgtagt gggaaagtta atcgtaggaa ttgggtcatt cttgtcatac 6180

ccaacagagc caagaaacca ggagggaaag acactcaggg tgcaaaatat tgtctgaaga 6240

atgtaattga aataggccct attatcccat ggaactaatg tttatggttt tttgaaggaa 6300

catagaaatt gactcctcca gtcttaaaaa ctcaagatag ttatattttt cttatctgag 6360

gagttctttt gtcaggaaac caaccatcag gcctccagat actatcaaaa ggagctgaaa 6420

cttacatatc actgaatcgg gacagtgaga catcagactc ttcacccatt gtgattgcct 6480

aactgacctc ctgcttcctg ttgaccaaat catcttcctt acccctccct aattcctgtt 6540

ttcccacatt tcttccctga tatatatacc cctcatttta gtagttcagg gagatacatt 6600

tgagaatggt gtcccatctc ctcggctgca gcacctgatt aaagcctgtt ccttggcaat 6660

acttgtctta gtgattgttt ttctgtgtgg tgagcagcag gatctacact gaatccctgg 6720

catttcagta acaaaattct ctgcaagctt cactgccttt ggcttattgt aacctgaaat 6780

caaatttatc cacaacttct gagataactt gatataactg taggattcac tttgtccacc 6840

actgcttccc agtctgagct tgccagctcc caacccttcc tagtgcccat gaactttctc 6900

aaagagccat aggtaacatg ttcccttttt cgtaaaactc taaccttctc tttgttcttc 6960

caacatattg aagaccactg agttttcctg tatgccccat ttggcaaata tttctttgca 7020

cgtaaaacat taaatttaga gattcatctc tacattttat ttagacttca gtagtttaga 7080

ctctaattgt ctgtattaag acaattcctg ctttgaatat ctatagtggc ctcttctctg 7140

ttatataaag tccagctgaa gccataaact agactcttca ggtgtcatga tctctgtctt 7200

tattaaatca ggagaggcat tgctagatct gtgcagttgg ggctgagaaa gagaaaagaa 7260

ttagggtgca gaggtgactc catgtccccc tctaccaaca ccatcagagt gtggctgcat 7320

ctgaggacca ctctcagctg atagaggcat caggaggagc agctggggca gccctgcctc 7380

acacatctgc ttccctgggg gtttatgttc gggtgtgtaa cactgtggga gaataactat 7440

tatactgttg gcagtaataa gttgcaaaat catcaggctg caggctgctg atggtgagag 7500

tgaattctgt cccagatcca ctgccgctga accttgatgg gaccccactt tctaaactag 7560

acgccttata gatcaggagc ttaggggctt tccctggttt ctgctgatac caggccaacc 7620

agctactaat actctgactg gcccggcaag tgatggtgac tctgtctcct acagatgcag 7680

acagggtgga aggagactgg gtcatctgga tgtcacattt ggcacctgag attggaaata 7740

gaaacacaaa tattcatact attgatcata ttataggaag acttccctga ataaccaggc 7800

agtactgagc acactgggct gagtaaattc ctagtgttct ccttccttac ctgggagcca 7860

gagcagcagg agccccagga gctgagcggg gaccctcatg tccatgctgt gtcctgactg 7920

ggtctgactc ctgcacaaag tgtgaccagc ctattaataa ggcttcaggg caggaggttg 7980

tgctctggga acatgcaaat gagcagggga tggggcaggc tgggcacagc tgcagagctg 8040

gctcatctca gtaactcagc accagctcag tgtccccagg tgtcccaggt aagaccaggg 8100

tagcacaaat ttgtctgcag agaatgtgtt tctactgggg actattttat tatgagaaac 8160

aatttttagg tatttttttg agaattttaa atattcctca ggagccgata gagtaatgta 8220

tttcattggt gtatcaggat tatttaggag aatattcttg tttgtaggaa acacatagta 8280

aaatgttaga tggtaggatt ctcaagtctt caaaagactc tcataagatt ccgggtaggg 8340

aagggggtaa ttgtgctata cctgcaacat ttctgtgagt ttaacattgt tcctttctaa 8400

aaaaaattaa aaataaaatg tatcggcatg atgctatata tttgtaagta ttaggtaatg 8460

gtgttatgcc tttgttctta ctagtattag atcaagcaat ttattacaga tatacaaaga 8520

tgataccgtg ttgtctccat gcatgcagca ctcacagatc caccactatc aagaactgca 8580

ggtctcttta atacccagag actaaatgag gtgcacctta ttcttgtttt gggtaccttc 8640

atagtctacc ttcttttctg ccattgggta ttatttccca aagttcatct gtcttagtga 8700

gggtggccac tgcacggagc atgtccctgc catgcaccat caatgacact ttcttcttat 8760

actttttatc agtgcatggg gacatcatcc tgacccagac accagcctcc ctgttaacac 8820

ctttaggaaa aacatactca atctcttatc aagcaattgt ctatgtacat ggagaaatca 8880

gttggatcca gatgaaactg gacatggatt tgcattcatt atatctcata tctctaatgt 8940

accctgaacg tcccagcctg actcagtagc aggggaagtg gatgtaacca catcagcatc 9000

agtgggctgc agcctcgggc tccacaaaat tttactgatg cctgactagg ggagcaaaat 9060

cacagtgctg cagcccatgc acaaacattt ctgctgcttt gtaagcagcc tgaattttaa 9120

gggaacttgc ttatattgga agaaaggaag aaagttccat ttgtcctcta aatgtttgct 9180

gaaaatgaac cgacaaaaga ataatgaata agagaaaagg caaacaaaat tcacttagag 9240

tgcggtggga tatcatagtg gggtgattac ccagataact caatgagatc cagttgttca 9300

tatttccttt ctagggaaga gggaattggg aagtgtaggc aacctggaga gaatagatga 9360

aaaaagaaat gcatcctcaa aagaacaggc aatagcctgc ctggataaag catcaacttg 9420

cagtctcttc tatttttgat tcatgttttg tgttaatctt ccctgatata aaatttccca 9480

ggaagaattt ccttgacaat tgtttttctt ctggagaatt tgcttttagg cagataaggg 9540

atatttagga aaagacttct tgtgcatttg ctgctttcta aatgcctttg gctttacata 9600

atcatcatac caatgcagca tagtttgaga tgttattttc tggattcctt tacttgcagc 9660

cacctgccaa gatcctgttt cagagagatg cagctgcaga ttgagtgagc agttgacccc 9720

tgaacaacat ggaggttggg gcactgacca caggtgcaga tgaaaacctg tgtagaagtt 9780

ttgcatttct aacttaagta ctaatagctt acttttgact ggaagcctta gtgataaaat 9840

aaatagttga ttacactttt ttacatttta tatatatttt atactctatt cttccaataa 9900

agtatggtaa agaaaaaaat gttcttaaga aaaccataaa aatgagaaaa tatatttact 9960

actgattaag tacttgcttt caggtgacac agaagaaaat ataagtgtat ctgcaaactt 10020

caaacccaag ttattcaagg gttaactgta ccatgatgaa tgtagcagtc cccatctgta 10080

gtctagggct ttctcctttg ctgtacctct gctcacttcc aatggccata tatatgtctt 10140

atgttcttta tgatcttggg cagagaggtc tgcctacatg cattgctggc cagatgtcct 10200

ggaatgtgta tctctgagga aagtgctatg gtttgactgt gtcctccaaa atccatctgt 10260

tgtaaagttc attctcagtg aaatggtttt tgccaggtgg gccctattgg gatgtgttta 10320

ggtcatgagg gtggagccct ctagtggaat acattaatgc cagtataaac agggtttata 10380

gggctggaat ctttctctct cttctgctgg tctgtcctga taagacatgg ccttccttcc 10440

attgaaggac tcaatgcccc aggcatcgtc ttgaaagcag agaaagctga ccttaacctg 10500

cccatgcctt gatctcaaac tttccgtact ccagaagtgt gagaaaatat atttctgttt 10560

tttatgaatt acacagcgac aaggaacctg ttatagcagc ttgaaagaga acaggagaga 10620

cagctcacaa tcagtgagga taagatgagg tatatacata tcccagcttt ctcatctctc 10680

aggtggaata gcccagagga atttagtcca tgtttccaca tgtggttgat cttcagttat 10740

cctgagtcag gtgggtggtt gatgtgtctt ttaccattca tcttctgctc ctgcctcact 10800

ttcttccttt cctcccagtg taaatttgct gcctaaacag gaatactcat tgctggtgga 10860

cccaaaataa gacagaaaaa aaaaatcatt gtacttttgt atgagggata tttctcatct 10920

gaattcttat cacttctctt tctttgacat ctaggaatat tcagaaaaca ctttttttta 10980

accaatctat tttagattga atttattgat tttttttcct gtgtgttttg tgaaccaaaa 11040

attaagttgt aagccaccaa cgagctaaat ggactcccct tttggcagag agaacttcaa 11100

agaaatctga aaaactaggt taggccatga ctggcaggtg ggtttggatg tgtctcatta 11160

tgctctcttc cctttggagt tcaggcacaa ctgaccagca ttatcattat aacagagatc 11220

tttggactga ggaaacagat gcttgtagca ataagatacc atactccaac atgacagata 11280

ataggccctg aagaaaatct aaaaatttta ctctaaaaat atttcttttt ctttttcttt 11340

tttttgtttt tttttgagat ggagtcgcgc tctgtgccca ggctggagag cagtggcgca 11400

atcttggctc actgcaagct ccgcctcctg agttcatgcc attctcctgc ctcagccttc 11460

caagcagctg gaactacagg tgcccaccac catgcccgac taattttttt tgtattttta 11520

gtagagatgg ggtttcaccc tgttagccag gatggtctcg atctcctgac ctcgtgatct 11580

acccacctcg gcctcccaaa gtgctgagat tacaggagtg agccaccgca cccaacctat 11640

ttctctgaaa tattctgaag tggccctgca aagctgcgcg ttgtgggaga aatttgcatt 11700

ctgtagagaa tctcctatgc ttactaatca ttttccaaag gtctgacttt ttttttttaa 11760

ggtctgacaa gcaacgtcta ctttttctgc tacccatagg attcatctac acgaaaagaa 11820

ccttggctta tctaaactca agcacacccc gttatctaaa ctcaagcatt tctttatggt 11880

gaattcaact ctttaggcag agcttaactc tttcaaccag ttgccaatca ggaaaacttt 11940

gaagccacct gtgacctgga agcccctgct tcaagatatc ccacctttcc agtacaaact 12000

aatgtatatc ttatatgtat tgatttatgt ctttgcctgt aatttctgtg tctccctaaa 12060

atgtataaaa ccaacgtgta atccaaccac cttgggcaca tgtttacagg accccctaag 12120

gctgtgtcac aggccataat ccttatcttt ggcaaaataa atctataact tgattgagac 12180

ctgtctcaga tactgttttg cttacactgg gtcacaaaat ttaaaaatcc tctaaaacta 12240

tctacacatc cataaatcta cattagaggc agtggagtga gcacactcca acagtcaaaa 12300

ctcacaagtt agaggaacct gagtctaaga ttttgtccag ctctcttgct tcataaaaat 12360

aatttgtgat ttttttaaat tttactctag agaaggacaa ttttggggaa tatgtttatg 12420

gtcagtggaa tgaataatgt ccccacccca aaagatgtcc atatcctcgt ctctggaacc 12480

catgttatac gagttattaa gaaactattt taggcatata gagaggaaac agggtccttg 12540

ggaaggtttt gtttctttta aagcagctcc agaaatgttt cttgtttagc aggaaagccc 12600

tggctcttag agctggtccg gcaagctttt ttttcttttt tagagaggga gtctcgctct 12660

gtagcccagg ctggagtgca gtggcacgat ctcggctcac tgcaagctcc gccctctggt 12720

ttcctcatgc ctcagcctcc caagtagctg ggactacagg cgcccgccac cacggccggc 12780

taattttttg tatttttcag tagagacggg gtttcactgt gttagccagg atggtctcga 12840

tctcctcacc tcctgatctg cccgcctcgg cctcccaaag tgctgggatt acaggcggca 12900

agctttgata tacaaatact ggccattaga aactgggtcc agccaaacat ggagattccc 12960

acattcttct tctttttttt tttttttttt tgagatggag tctcgctctg tcgcccaggc 13020

tggagtgcag tggaacgatc tcggctgact gcaacctctg cctcctgggt tcaagcgatt 13080

cttctgcctc agtctcctga gtagctggga ttacaggcac atgccaccac gcccagctac 13140

tttttgtgtt tttagtagag acggggtttc accatattgg ccaggatggt ctccatctct 13200

tgacctcatg atccaccttc cttggcctcc caaagtgctg agattacagg tgtgagccac 13260

ggcgtccagc tcccacggtc ttcttccttg ccccgacatt tgcctgacaa catggctgcc 13320

cccacatatc cccatgtgtg tagaacttta tggcaccctg catttgcata ttaaaacact 13380

agggtgggag ggccagtttt ttctcaagct acatgaatga catgcttggt aaaaccaatc 13440

ccctaagccc tatgcaaatc agacaccacc tccttcagcc tcctcatata agcagccact 13500

tttccaccgc acatggagtt ttctcttagt tctaatctcc cctctctctg tctctgtacg 13560

gtggagctgt tttcttcttc cttccttctt gcgtattaaa cttttcgctc cttaaaacaa 13620

ccccacgcat gtccgtttct ttttaaacaa acccgcgtga gactaagaac ggtggtggtc 13680

ctccagtcat cagagcccta tcataaggat tctaccttac atagcaaaag aagaacttga 13740

caggtgtaag ttaacgactt tgagaataag caacctggtt atccagatga aaccaatata 13800

atcacaaagg tcattaaaat agaggaggag gcttgcagag aggagctggg acaatatgga 13860

aggatgtggt ttgaggaggg aaggggccat agagccagaa atgtgggagc acctggaaga 13920

taaaagcaga ggattcagtt ctttcctctg gagcctccag aaggaatgca gccctactga 13980

caccttaaca ttagctcagt gagaatgctg acctccagaa ctacagggta atacatttat 14040

gttgtgtgaa gccaataaga ctgtggcgat ttaaacagca gcattggaaa ctaatgcaag 14100

gggaagaaat gtctttcagc tcactgagcg cgtgcttgtc ttctgttctt ggaaatattt 14160

ccaccttgtt ttctggtgtc agttatgaca agagacagag aaaatttttc cgagagaaga 14220

gctaccacga gaattcgttt ttagctagaa aatctcctgg gtaaaatctc atgattatct 14280

gttatacgat ctgggtatca cagagtttgg ggtcagatct cacaacatgt gataggagga 14340

agagggtttt gttagtttgt tttcatattg cgagggaaaa ttagattttc aggacacaat 14400

ctgagaagga cagacagagt cagagatatt gtaagagaaa gaggagatgt ggaaggagtc 14460

agggcaatga aacacgtgtc ccagctccca aatctaaaat aaagtcattg attttacaag 14520

gtgaagaaaa gctcttcacc ttccatcttc ttaggaagga tcaaattctt cccagagtcc 14580

ctgcccctcc ttcacccccc tgacattgca ttgtggagct aatcacatgt gcccttaatc 14640

cctgctcctg tcccaggctg ggagaggctc actgtcctca ccatgcccag caggctagag 14700

ctggtgccct aagcaccaaa ggagaagagg atgatttctg tgcaggcaat aagagcttta 14760

ccgcatttta catttcccta aaattctaag caaagttctg tggagagaag agaacatggg 14820

gcaactttat ctggtggggc tggaagagac caggctctga gaatgacaat taattcacat 14880

ttctactttt ccagatgcca aatgctagct ttatcaattg tggaaatgtg gactttgagt 14940

ttaagaacag gtttcaggta ctgcaataat tgaggagaca tttggaatag agtccttagt 15000

ctagggctaa agtcagactg gcagtgcctg agagcttcca ggcgctggcc ctgggctgtg 15060

agggaagcag cagttctcct gagccacagg gctgaggacc tgggagcaac cacaggcctt 15120

ggccacaggg ctgcctggca ggggcttgag ggaaggaaac tgctcacaaa ttcacaggag 15180

ctgcattaag catatatccc gcagcctggc aagagtgaaa gtctcagaga cccagacctt 15240

agggctgggg ctgggattct gggctggctg ctgtcggctc tgccctccct tgtcctgaat 15300

gactggcacc ctgctggagc caaaaatgga gggtcggcca atgtcctcaa agtctttact 15360

cagccagacc ctgttctgct tggaaagaaa atgaacacat ggtatacaaa taaacataaa 15420

atcatgtgta tctgtaataa gaatttctaa tataaatttt acatattgaa tacatatatt 15480

ttagaaataa gttgtactta tacatttgcc tatactatgt gtctatttat ggataaatat 15540

gtggtacatg tttatatata aagatatata actttggttg ctagtataag gtatagtttt 15600

aaactttaat aaagtaaaca cgaacattga aaatgagcac tgggtgcacc acttcatggc 15660

cctcctcact ccagggccta agggtgataa agctcaggac cctcttccat gtcccccggc 15720

atggacagca gtgaactacc cattgctgag gatcatgagt gacctgaggg ggatctgcag 15780

aatcacagag cagcagatgc ccagaagatt gatagtttgg accctgaatg gagggacatt 15840

ttgatcacag ggctcagcat agacaagtac tcttctggtc atttctgtca aacaaaacag 15900

cacatgtagc tcagaggctg catgatcaca ggatgaagca cctccatgag tcattgccag 15960

cttcccctct gactggggtg atgtgggatc tctctgcccc actttcatgg ctcaccagct 16020

gctgggactc tgttgacagt ggtctacatc tgtcccacgc acaaacttct caatagttgt 16080

cattttactt actggtctcc aataaccaca aattgccagc catgaccctt ccacaaaaat 16140

tgatggagtg gggttggaac tatgggttca gcaatgtgcc agggcatgta tataagagaa 16200

acaaggtgtg gccagcccat gtctgaagtg aggatgaatt ttcaccatga attcggagta 16260

ggaggtggaa gaaaacatcc tgacatgctc caggcagtga tgcccacaag atacactaga 16320

agcgcccaca gaggaggtgg gaagggggtg tgagaaaaag agggagtgtg aatttacagg 16380

tgtctccaga ctgttcacct gaaaagagat cagaccctta acaacacctg ggaccccagg 16440

cagagctgtt gctatggaag atttgtgcag gacagggata ggaggaggga gcacctacct 16500

gtgatggcat cttggttgct gttggccaga atgacatctt ttcaggccta ctgtccctga 16560

cccaggcagg gatcatgtcc aggagggcag caggagctca gagccaggcc ctggttctgc 16620

tagtgccagg ctaggatgtt cttcatactc tggccagctc tacaggtgac agtgaccctc 16680

tcccctgaga cacaaggagg aggacaacga tggcatccac ataacatgct cactgtcatc 16740

caagtggggg aacaaacatg cagatcccca aatattaata ccgagtttgt agttcctcca 16800

acttggtgca attctgatca gaaggaaaaa caggctaaca acttcatcag cagggaaagt 16860

tcatgtttaa tacaaactat ctgaggctga agcctgagtt tcccttcctc accaggcagt 16920

caggacagca ggagcaaaag gagaaaagct gggtcccaaa gtccacaagg tgcctcctga 16980

gactgatcct gcttagagaa ggtggtgaca gtggatgagt cttctgggac tcccacacca 17040

acataccctg agctggatgg cttgagccat agacatttat tgctaattta ttaaacgttt 17100

attgtgaatg tattagaacc ctggaaagtt caagatcaaa gtccagaagg atttgctttc 17160

tggtgagaac cctgcttctt gttttcagat gtcacccctg gatacatcct cacgtgtgtt 17220

cagcaatgtg ccgggcatgt gtaagtagga gaagcaagta catgctttct tcagtgcatg 17280

cttggaggct ggtgaggggt taaggaacag atcgatgatg tttattttta ctaggacaca 17340

aatcctatta aatcagagcc ccaaggtttt cacttcattt caccctaatt accccttcat 17400

aacctctatg tctaaataca gtaattttgc aggttggaga ttcaaaatat gaatttgtga 17460

aacacaattc agttcatagc aggtgtgctt tgaggacatg cccaaggcag taggaagggc 17520

aaggcgaggc ttccagtctc agagcacaga aggcgttttc ccaccactca gcacactggc 17580

agctcctctt aggtgatcca ggtcacacat gagacgccat tcccgccttg ggggctgcct 17640

gctgataatg gaacaatatc acccttaatt ttcactattt ctaagatcag acttctattt 17700

tatgtttatt atggagtttg tccaatggcc aggtttgtaa cataacctcc aatgtttggt 17760

ttgtgacata acatcttata gaattttaga tgctggtttc cagtgagact tgatggagtt 17820

ttttcatggg ttttcatgga gttgatctta ttacatccac cattatgctg cattctgtag 17880

ggaacacaaa atgattcctc tcctcaataa accagtcacc tagttgaaga taaagggtgt 17940

tcagaaaaga tggtcacaaa acaattaggg gctagattct gtgatccatg tgagatgcca 18000

atgatgtgat tctggaaagc agaaggtgag gttgtgagat accctgcagc acataatgcc 18060

caaggctagg gtggcgagta gagggcccct ggtctggatc tgtctaccca gcttcattcc 18120

tcccattcgt gaaacagtgg ctgcaccagc caggaagcat ctgagaccac ctgattctct 18180

ggaataatag ctgctgaggt ttgtgctgtg agctataaca ctgtgagtct ctaactgtaa 18240

ccatttagat tagcactaat gaaaataatt tttaagatgg tgaaaatgtc tatatctatg 18300

ctttgcaagt gtagccgtta tatgtcatat gtgactcctg aacacttgaa atgtagtgaa 18360

aacaactaag gaactatatt gttaatttta attaattcca atatacatgt ttgtggctgg 18420

gtactgctaa tgttggccat gtgagtcagg gaagttttag atggtaataa acagcaatgt 18480

catcaggaca accaatctgc taaatgttaa gcaaagatga tcccaggacc attgcaccaa 18540

gctggagtgg acaaggtgga aagagcacgt aggtggatct ttggaaccca gattctgctt 18600

cctccaggtg aatgatgtaa actctctgtc ttgaagacac tgacattgaa gctagcgact 18660

ggggagctga gtgtttctcc tggaagctgc agtggaaaca gaacatagat cttccttagt 18720

attactcatg tccctctcca taggcttccc agcagagagg tgctgtctga tcaagtggga 18780

ggtgaatgct gtgctcccta atcaatagtc tttctcagtc ccatgggaca tatgaaccct 18840

gtcacatgca gcccacgcag ttgaagccag agccccatgt ccagtgtcca gtgttgcctc 18900

tgccttcggt gatcccaccc tacaggaccc agtgatgaag gcttgagtgg cagggaagag 18960

ctttggggag cctgccaact agcaataagg gtgcagactt gggcttccag cttctgagca 19020

agggaacatt gtatgtccta ggtgtgatcc aggtcaaggt gtttgataaa agatattaca 19080

ataaattatt ttcttaagaa aataaaaaca cagaaggcat ctgattatta taggactgtt 19140

tgggttttat accactaaag tggtaaaaat aaatttttca atgcgcagtg gtttatcagg 19200

aatctcccag cccctgccct tcccgtgatg attctgtagt ctttgcaggt tgtatggtca 19260

cttggtggcc tccttaccga tcagatgcat ctacagcaca accctaagat cttccccgat 19320

taacatagat gagcaacttc aacctccaga taccacatac cattggctga tttttatgca 19380

atttatcaac actgaaagta ttctattcca tttcattatg tagttattat tctgtgcatg 19440

gctcattccc ctactgagta tcagtatttg cgatcaaaaa ttgttttatt tacttacatg 19500

cactaaacct taaatgatag catttattag ttctgaaaac tctgacatta agttaaaatc 19560

ccttaaaagc aactatatgc ttcttttttt aattagaata aatatggtgc aatcacctca 19620

actaaggata ctaaactact tttacagttg aaaatgatta attggacatg gggaaacata 19680

ttgacaaaag taacaagcat tattatgatg ttgaataaac acagtttgca tgtaaagtgt 19740

acagttatga ttaattgtat taaaacacag gaggaagaaa ggtggggatt gctggattgg 19800

ttggtgattc agtccttaga agcaaggggg agctgatgag tgatctcagg caggcatcca 19860

atataaggtc caaatatgat taattacaat gttcttttcc ttaaggtttg atattttaag 19920

ctggtcctgc tggggaagtt ttgagcacat tcatggaata tgtggaataa agattaaaaa 19980

cagtatattt caaaatactc ttaaaaagct cagaaacacc caaagaaaac tattttataa 20040

aatatcatgc cttcctcgcc cattcttgat atttgtgttt aaccaagcat gactcacaat 20100

tcaacttttc tccacattgt tgcctagagt gttgaagaaa ttcctctaat aggacagaaa 20160

tggagtcaat cttgcaatat ctggccaatc gtgttctggc cagtcttgct ttatctcaga 20220

atgttacatt tccagaaact tctacagtta ttcctgaaaa ctacaagcaa gaaagggaag 20280

gagactggga tcattcaagg caaaatggaa cctgttctgc aaatactgta gtgaaatcat 20340

aactcagtaa tcgttttgtg actggcttat ttcacttagt gtaatgtcct ctagtttcat 20400

ccaagttgta gcatgtgtca gaatttccct ttttaaagct aattttgcct ttttaaagct 20460

acaatatgct actgtatgta tataccacat ttggattacc agttccctcc tttgtgaaca 20520

tttgagttgc ttctaccttt tggctattgt gaataattcg gttctgaatg tggatataca 20580

aatatatcct caagttaatg tcttcagtta cttgggtata tgtccaaaag tggaattgct 20640

gaattatata gtatttctat ttttaattct tcgaggaatt gccatctggt ttcccacagc 20700

aggtgggcca tttacatcac cacgacagtg tccacaggag ttccagttcc ctaagttctc 20760

accaagactg gtcattttct gttggaaaaa aaatcctaat aggtgtgagg tgggttttgt 20820

ttttattttt ctaaggatta ataatgttga ccatgttttc ctatgctagt tttctaatta 20880

tctctataga atcttcttta gagaaatgtc gattcatgta ctttcctcat ttttaatcag 20940

gtattttatt ttaaggttca catataagac ttatttttgt atagtagata ttattaaccc 21000

cttatcaaat ataagattta caaatgtttt cttctattcc acatgttgca ttttcactgt 21060

gttgattatg tcttttgatg cccattttac atttttatga agtccaattt atcttctttt 21120

ctacttttgc ctgtattttg gtattaaagt tgttagtatt taaatgtcta taaatactga 21180

cttactatgc tgagaaggtc acactgcacc atctctctga tggtggagct aagagtttta 21240

cactctccca ttttgtaccg ggggacagtg cagctatgtg agaacccagt ggctttaccc 21300

agcttatttg tcttgcattt ttggtgacat ggattgttat tcacattggc ttcctctggc 21360

aggtccacct ggaaagcatt atatttagca agaaaaaagg aggcagtgaa tgatgtcact 21420

gtggactgtg tatattccct gttgcaaatg tcaaattcgt gaagcataaa gggatttact 21480

agaggatatt aaattccttg caaattgttg aaaagcctta aggaacaggc tccaggcaaa 21540

gcctctagaa caattccaag aatggcactg ctggcacagg ctggggagga gctcctcctg 21600

cctgagactc cacaacattc aagctgtctc ctgcagaaaa gagagcagct cttctcacta 21660

ctgcccccag aaggacagcc gctctgctat caactacgag agatctgacc cctttctctg 21720

actgcccatc agatgtgtta ctacagagtg cttttggatt gagatgaggg agggaaaatg 21780

cctgatctca ctcaaatgtg ctcattttcc tggtctgggt ctcagccagg cagcacagtc 21840

ctgtgggcac agtatcttca cttgccaggt tcctataaaa tgagagtatc tttcttgact 21900

gtggaaaata tggggatggt aagggtcaag gaataggaaa aggagtactt ggagttgcct 21960

gagctgccac tctggtttcc ccaaatgttt tcaatccatt aaattcatga taataaacac 22020

ctttctgcct aaccagccat aatcgctttt cttagatcca aatgattctg tgataaaact 22080

aaagagaaaa aataaataca cttttcaacg tttaagctga ccaggaggct gtctctggtg 22140

tttacatgtc catatcccac tatgcatttc gtcttccctt tgcctcactg gagatccaag 22200

tgcagggaat gcttcatatg actgtgatcc ctagaacttc atcctcatga ctgttgaaat 22260

cttctctgag ttttaccagt ggaaatggca ataaatagaa agattccaga aggtccccta 22320

gggtccatat tttctgtact tcctctctaa aatatgtaaa atcaattata tttgctcttg 22380

ctttatatgg gctacatctg tgtccccagc acacacccta acgccctgcg ttgacaactt 22440

atccagtatt gtattaagtc taaataggca catgtttatc atattatttc ttccaattta 22500

atgagataat tgtcatgtca aatctttgga ttaagaacca tcagccagaa aaccacagcc 22560

ctgagaaaat agaaatagag tatttcaagc tgttcagttt gcacaacaat gatgattcta 22620

tagctcattt acaagatata aatattctgt ttatgggagt aagagcaaca ttcacggtga 22680

gtgttgtaga ccctccgcca catctgacag catagcaatt caatagcaca agggtttgtg 22740

tgccagcagt gcttgactaa taatgggttt tttttctata gggaaagctg cacctgtgca 22800

catgacagga tcatgagcta cactttactt cacttttttt gaagtcaatt accttcttga 22860

tccaaagcca cagtatggaa gaacttcctg actctgatga aggcatttgg aaaatcctca 22920

aataatattt tatgcagtaa cataaccaaa gacaaaggaa aattcatata taggataata 22980

accactgctc tcctcatgtt acatgaagtg cacttgaatc acctgacatc agctgttagt 23040

atgagccctg atgtgtggta gaatgttagg ggctcaggaa gggtcaattt tcttggcaaa 23100

ttagtgttca agagagccag tagccctgta catctcagtt agttgaaact catattattg 23160

agtccccaca gggacctccg taacagtggt tcatgaggtt acctgggaaa gtgggcaaag 23220

gtgactgact gaagtccatg tgttcaatca tcctcattat gaaaagcccc ctgctcatta 23280

atggcatttg gattaatatt cactgagaaa gcatgttttt cgattatggc tcatttttga 23340

aacatctagt catagctctt ccaaaatgac tttgtctgca atcatcctgt tgtgtttctt 23400

ttaaggcctg atgatctgtc aaaaccaaca accaatatta ttaaattgct gttctctttg 23460

caatgtggca atgtgatcat cagatgtagg ccttgcagtt tacccacggt taggattttt 23520

acatttccag ggattccttg acaatggcac taatgtgacg ccatctagca tccattgttg 23580

tgagaactct atttggaata tcatcattgt gggcatgaac ataatggaga ctttcatagg 23640

atgcaatgat taatattcaa cattagtcaa aatctgctgt agtcctaccc ttggagatct 23700

actggtttgg aaaaatattt taggtaagga caacaatttg gtaaagtttt aaaaataaag 23760

gctaaaaaga aattttcttt agcaagagta tgaactatct gtggtccaga ataccatttt 23820

tttacagatg caagaaaaaa ttatttaaat gtatataatt tagataataa ttttgtaaat 23880

ttgtatatat tgcatatata atagaataaa cataattaaa tatgaatact atatatactt 23940

atagccaaca ttttatcaaa taaatatata ttaacttcta tgtatcttat atatatgcac 24000

atttttccat aaataaggta aattttaatt ttttaatgaa atactaatta gttttaattt 24060

catctttaat tttctcttaa taacttcata atttgattac ttaatgttca catttatacc 24120

accaatttaa gaaaaaacac atgtatctat atataggcag aggtgtaaat actgtatcag 24180

taagctttta tgcattaatg attgataatc ggttaaatat acaactcagt ggctgataac 24240

tgtaaatatt tgttttcatc ttcatggatg ctcatgttca caatcatctg gctgattaag 24300

gaagggctca gctgattgat tcctccgcag ggcactgagc ttgtctttag cctacagata 24360

ccaattttct gaggacgagg ctgaacagca gtgactacac gtgacacaag attcttgtga 24420

caggtcacag gagtgaacaa catcccaaac cgaactgcac agttgaattt aagtccaata 24480

atttctaaca tagcttcaca cattttaaat atattttttg atttcagtga gtacaaattt 24540

tcaagaaaat gtttactcca attaattata gaggtgtttg atcattccat ggacaagtaa 24600

ttatgttttc aacctttaca atcctgaaaa tatttgcaaa cgtaaatttg cattaataag 24660

aaaataaagc tggatgtgtt ttcaacatgt gcttttaaat ataatttttg aaatgacctc 24720

aaagggggaa aatcatttta acttatgtaa ctatcccttt ttacctctct tgtgtcctat 24780

agagttgccc aataagaggt cctcccatga gatttggaat cagaaaagga tgactcaata 24840

ttctccattg gtacctaagg cagacacagg aacagaggtg aggactagag aaacacctga 24900

aaagatgctg taggaagctg agagcatcag cacccccacc cctaagcttc cagacaggac 24960

tgaggaccac atggttagaa agcccttact tcagggacag atgcattctg ttttctgagg 25020

gagcaccaga gattcctgct tctaatatca gctttcctga ctactatatc cttggctttg 25080

aaaggtcata gtgggaaagt taatcatagg aattcggtca ttcttgtgat acccgacaga 25140

gccaagaaac caggagggaa aggcactcag ggtgaaaaat actgtttcta gaatgcaatt 25200

gaaataggcc ctattatccc atggaactaa agtttatggt tttttgaata aacagaaatc 25260

gactcctcca gtcttaaaac tcaagatagc aacatttatc ttacctttct tttttttttt 25320

tttttttttt taaatttcag gaaagcaacc atcaggcctc ccagatacta tcaatttgct 25380

gaaacttata tatcactgaa tcgggacagt gagacatcag acccttcacc cattatgatt 25440

gcctaactga cctcctgctt cctgttgacc caattatctt tcttaaccct ccctaactcc 25500

tgtttttcca catttcttcc ttgttatata aacccctaat ttcagttggt cagagagata 25560

catttgagaa tggcatccca tctcctcagc tgcagcacct gattaaagcc tgttccttga 25620

aaatacttgt cttagtgatt ggctttctgt gtgacgagct gcaggatcta caccgaatcc 25680

ctggcatttc agtaacaaaa ttctctgcaa gcttcaccgt tttggtttat tgtaacctga 25740

aatcaaattt atccaaaact tctgagataa cttgatataa ttctaggatt cactttgtcc 25800

accactgctt accagtctga gcttgccagc tcccaaccct tcctagtgcc aatgagcttt 25860

ctcaaaagag ccataggtaa cattttccct ttttcataaa atgctaaatt tctctttgtt 25920

cttccaacat attgaagacc actgagtttt cctgtatgcc ccatttggca aatatttctt 25980

tgcaaataaa acattaaatt tagagattca tctctacatt ttatttagac ttcagtagtt 26040

tactctaatt ctctgtatta agactatatc tgcttcaaat atctatagtg gcttcttctc 26100

tgttatacga attccaactg aagccataaa ctagactctt caggtgtcat gatctctgtc 26160

tttattaaat caagagaggc attgctagaa ctgtgcagtt ggggctgaga aagagaaaag 26220

aattagggtg cagaggtgac ttcgtgtccc cctctaccaa caccatcaga gtgtggctgc 26280

atctgaggaa caatctcagc caatggaggc atcaggagga gcagctgggg cagcccagtc 26340

tcacacatct gcttccctgg gggtttctgt tcgggtgtgt aacactgtgg gagggtaatt 26400

gtaatcttgt agacagtaat aagttgcaaa atcttcaggc tgcaggctgc tgatggtgag 26460

agtgaaatct gtgccagatc cactgccgct gaaccttgat gggaccccac tttgtaaact 26520

ggatgcagca tagatcagga gcttaggggc tttccctggt ttctgctgat accagcctaa 26580

atcatttcta atgccctgac ttgcccggca agtgatggtg actctgtctc caacagatgc 26640

agacagggag gatggagact gggtcatctg gatggcacat ctggcacctg agattggaaa 26700

cacaaaaaca aatggtccac acaattaatc atgtagtaag agaatttccc tgaatagcca 26760

ggctgtgctg agcaccctgg gctgagtaaa ctgccagtgt tctccatcct tacctgggag 26820

ccagagcagc aggagcccca ggagctgagc ggggaccctc atgtccatgc tgtgtcctga 26880

gtgggtctga ctcctgcaca gggtgtgatc agcctgttaa taagtcttca ggtcaggaga 26940

ctgtgctctg ggaacatgca aatgagcagg ggaaggggca ggctgggcac agctgcaggg 27000

ctggctcatc tcagtaactc agcaccggct cagtgtcccc aggtgtccca ggtaagacca 27060

gggtagcaca aatttgtctg cagagaatgt gtttctactg gggactattt tgttatgaga 27120

aacattttaa agatattttt tgacaatatt cctcgagagt caatggggta atatatttca 27180

ttggtgtatg gggattattt tggagaatat tcttgtttgt aggaaacaca gtacatatta 27240

gatggtacga ttctcaggtc ttcaaaagac tgttataaga ttccatttag ggaagggggt 27300

aattgtgcta tacttgaaac atttctgtga gtttaacatt gttcctttct aaaaaattaa 27360

aaataaaatt tattgacatg atgctatata tatttgtaag tattaggtaa tggtgttatg 27420

ccattgttct taccagtata agatcaaaca atttactaca gatacacaaa gatgatgccg 27480

tgcttcttca atgcatgtgg cactaacagc cccaccatta tcaagagcta caggtctttt 27540

taatacccag agactaaatg ggctgcacct gtgtagaatt tttgcatccc taacttaagt 27600

actaatagat tgcttttgat tggaagcctt agcaataaaa taaatagttg attaacatgt 27660

tttttaatgt tatatatatt atatactcta ttattccaat aaagtatgct aaagaaaaaa 27720

atgttattaa ggaaaccata aaatagagaa aatatattta ctacttatta agcatttgct 27780

tacaggtgac acacacagaa caaaatataa gtgtatctgc aaatttcaaa cccaagttat 27840

tcaagggtta actgtaccat gatgaacgta gcagtcccca tctgtaatct agggctttcc 27900

cctttgctgt acctctactc atttccaatg accatatata tgtcttatgt tctttatgat 27960

cttgggcaga gaactctgcc tggatgcatt gctggccaga tggcctggaa tgtgtatctc 28020

ttaggaaagt gctatggttt gactgtgtcc tccagaatcc atctattgta aagttaattc 28080

tcagtgtaat ggtttttgcc aggtggggcc tattgggatg tgtttaggtc atgaggctgg 28140

agccctctag cggaatacat taatgccact ataaacagga attacagggc tgagatctct 28200

ctcctgctcc tctgacatgt taagacatgg ccttccttcc tttgaagaac tcaacgctcc 28260

aggcatcatc ttgaaagcgg agaaagcaga ccatgtggtt gatcttcagt tatcctgagt 28320

caggtaggtt gttgatgtgt cttttaccat tcattttctg ttccctccct cactttcctc 28380

ctttcctccc agtataaatt tgctacctaa caggaatcct tattgctggt ggatccaaac 28440

tgagagagta aaaaagaaaa gtcattaatc ttttgtatga ggggtatttc tcatctgaaa 28500

tcttaccatt tctctttctt tgatatgtac aaatattcag gaagcaccta attttttttt 28560

ttttaccaat gtatcttaga ttgaatttac gctgtgattt tttttctgtg tgttaaaaca 28620

aaaaatcaag ttgtaagcca ccaacctact caatgggctt ctcttttggc agagagaacg 28680

tcaaagaaat ctgaaaaact aggttaggcc atgactggca ggtgggttta gatgtaactc 28740

attatactct cctccctttg gagttcagac aaaactgacc agtgttatca ttacaacaga 28800

gatctttaga ctgacaaatc aaatgctttg tagcaataag ataccatact ccaacatgac 28860

agataatagg ccctgaagaa aatctaaata ttgtacccta aaaatatttt ttttgatgta 28920

ttctgaagtg gccctgcaaa gctgcctgtt atgggggaaa tttgcattct gcagagcatc 28980

tcctcctctt actatgtctt ttccaaagag ttggacattt ctttaaaggt ctgataagca 29040

acattcacca tctactttac tgttacctgc aaggttcatc taagtgacaa gaaccttgac 29100

ttccacaccc ccttatctaa actcaaggat ttctttatga tgaattcaac tctttaggca 29160

gagcttaact ctttcaacca gttgccaatc aggaaaactt tgaagccacc tgtgacctgg 29220

aagcccctgc ttcaagatat cccacctttc caggacaaac taatgtatat cttatatgta 29280

ctgacttagg tctttccctg taatttttgt gtctccctaa aatgtataaa accaacgtgt 29340

aatccagcca ccttgggcac atatttgcag ggcctcctaa agctgtgtca caggccataa 29400

gccttatctt tggcaaaata aacctataca ttgattgaga cctgtctcag atactgtttt 29460

gtttacactg ggtcacaaaa gttaaaaatc ctctaaaact ctctacacat ctataaatct 29520

acattagaga cagtggagtg agtatacctc aactcaagtc atactttgag tttaagaacc 29580

agacagattt tcaggtactt cgataaatga ggagacattt ggaatagggt ccttagccca 29640

gagctaaggt caggctggca gtgcctgagc gcttccaggc cctggccctg gtctgtggag 29700

gaagccactg ttctcctgag ccacagggct gagaacctgg gatgagccac aggccttggc 29760

cgcagggctg cctggcaggg tcttcaggga gggaaactgc tcacaaattc gcgggagctg 29820

cattaagcat atatcccccc agcctggcaa gagtgaaagt ctcaggacac agaccttagg 29880

gctggggctg ggatcctggg ctggctgctg tcagctgtgt cctcccttgt cctgaatgac 29940

tgggaccctg ctggagccaa agagggaggg tcagcaaatg tcctcaaggt cttcactcag 30000

ccagactctg ttctgcttgg aaagaaaagg aacacatgct acaaaaataa acataaaatt 30060

atgtgtattt gtaatatgaa tttctagtat aaactttaca tattaaatac atacatttta 30120

gaaataagtt atatttatat atttgtatat actatgtatc tgtggataaa tatgtgatgc 30180

atgtttatgt ataaagagat gcaagtttca ttgctagtat aaggtatagt tttaaacttt 30240

aataaattga acgtgaacat tgaaaacaag cactgggtgc accacctcat ggcccctcct 30300

cactccaggg cctgagggtc ataaagctcg ggactctctt ccatgtgccc cctgggccga 30360

tagcagtgag ctacccattg ctgaggatca tgagtgacct gagggggatc tgcagaatca 30420

cagggcagca gatgcccagg agatggatag tgaatttgga ccctgaatgg agggacgttt 30480

tgtccacagg gctcagcaca gacaagtcct tttctggtca tttctgtcaa gcaaagcagc 30540

acacatggct cagaggctgc atggtcagag gaataagccc ctccatggct cattcccagc 30600

ttcccctctg actggggtga tgtgggatct ctctgcccag ctttcatgac tgaccagctg 30660

ctgggaccct gttaacaatg gcctatgtca gtgccatgca caagcttctt gatagttacc 30720

attttacttt ctggtctcca ataaccacaa attgccaacc atgaccgttt cacaaagatt 30780

catggagagg ggtcagaatt atgggttgag caacgtgccg gggcatgtat gtaagagaaa 30840

caaggtgtgg ccagcccatg tggaagtgag gatgaatttt catcatgaat tcagagaagg 30900

aggtgggaga aagcatctga catgcttcag gcagtgatgg ccacaagata cattagagat 30960

gttcatagaa gaggtggcaa gtgggtttga gatgaaaagg gaatgtgaat tcagaggtgt 31020

cccccagcct gttcccctga gaagatttca gacccttagc aacccctggg cccccaggta 31080

gagctgttgc tagggaagat ttgtacatga taggggacag gaagagggat cacctacctc 31140

tgaaggcatc ttggttggta gtggccagag tgacatctgt tcaggcccat tattatccct 31200

gacccaggca gggatcatgt ccaggagggc agtaggagcc agcaggagct cagagccatg 31260

ccccagttct gctggtgcca ggctaggatg ttcttcatac tctggccagc cctgcaggtg 31320

acagtgaccc tctaacctga gagacaagga agaggatgaa gatggcatcc acataacatg 31380

ctcactgtca tccaagtggg ggaacaagca tgcagattcc caaatattaa taccaagctt 31440

ttatttcatc caacttgatg aaattctgat cagaaggaga aacagtctaa caaattcatc 31500

atcaaggaaa gttcatgttt aatacaaact atctgaggct gaaacctgac ttccccttcc 31560

tcaccaggca gtcaggacag caggagcaaa aggagaaaag ctgggtccca aagtccacaa 31620

ggtgactcct ggggctgatc ctgctcagag aacgtggcaa cagtggataa gtctgccgag 31680

aatgccacac catcatactc tgagctggag ggcttcaacc acagacattt attgctaatt 31740

tattagacat ttattgcgaa tatattagaa ccatggaagg ttcaagatca aagtccagaa 31800

ggatttgctt tctggtgaga accctgtttg tttccagatg tcctctctgg atacatcctc 31860

atgtgtgttc agcaatgtgc caggcatgtg taagagaagc aagtgcatgc tttactcagt 31920

acacgcttgg aggctggtga ggggttcagt aacagctcaa tgatatttct ttttattagg 31980

acacaaatcc tattaaatca gaaccccgat gttttcacct catttcaccc taattacctc 32040

ttcataacct ctatgtctaa atgcagtaat attggaggtt ggagattcaa aatatgaatt 32100

tgtgaaacac cattcagttc atagcaggtg cactttgagg atatggccaa ggcagtagga 32160

agggcaaggc aaggcttcca gtctcagagc acagagggca ttttcccacc actcagcaca 32220

ctggcagctc 32230

<210> 81

<211> 116

<212> PRT

<213> Intelligent people

<400> 81

Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp

1 5 10 15

Phe Pro Gly Ser Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser

20 25 30

Val Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser

35 40 45

Gln Gly Ile Ser Ser Trp Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys

50 55 60

Ala Pro Lys Leu Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val

65 70 75 80

Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

85 90 95

Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln

100 105 110

Ala Asn Ser Phe

115

<210> 82

<211> 32230

<212> DNA

<213> Intelligent people

<400> 82

gaattcacca ttttaaagtg tacaattgag tggcatttag tacattcacg atgttgtgca 60

atcatcactc ctattattac tctgtaaact ttacaaattc atcgtattta tttatttaat 120

ggttgttaca gtttgcaact aaaatagaag gaaatacatc aaaatattag cagtggctct 180

tctgggtgaa gtacaagggg caaattgttt tctcctgttt gtttttctgt acacgttgtt 240

tactgtgtca cattgtttac tgtgaatagt tactatttga aactaaagat caagggcgat 300

aaactgtatt ttaagaggaa gtcagaaaac aaacaaggtg gccctctcgt atgctgtgct 360

gacacctgtg tccctgtagc ctcaactacg cctgttgccc ccacctgcgc ctgaagtttg 420

aggtcgctgg tggcacctct gtgccagggg gccctggaac ctctcctagg atggtcccag 480

agaaaggaga gagatctgct tcctggaatc tcagctctag atgctttggt caactccatg 540

tcatctgtgt ttgataagct tgatttccat ttgttttggc ccctctttta gtgtgctttt 600

tgttagcaga aactcacgaa ctagagaact gcaggtcccc caggggagtg ggttgagatg 660

tttttgacat cagccacctc tttctcccac agttttgtta ttaaaacccc attttaacaa 720

ccttgaaaga caggttttaa aagaaagttg caattttctt acaattctac tgcttaatta 780

aaagagtagg ccctgtggtt ccctttcccc atcctggtgg tgttctccct gcccctgtgg 840

tggtgttctc cctgcccctg cctctcacca gtgtatgctg ttcccaggtg gcagtaatcc 900

tatctcaggt ctggcattcc aaggcaggct cctgctcctg ggtggggagg ggcctccatg 960

ctgggttgaa tgtctgtgct ttgctgtccc ctgacccaca gagtgccctt ggacaggaca 1020

taggtctcag tttcctaagt gagagtcccg gattagatgg actgtggtta accctttatg 1080

tgtcatggcc tctttctgta taaagccaca gatctttccc tgcaatgagc atttatacag 1140

acacttttca gggttttgtg aacccgtgag ctggacatcc tagagctctc cttcaatccc 1200

atggcctcta gggtaccctg ctctggaaag gcaagactgg gcctcttttt tgggaccccg 1260

gagggcagac cttgggtttc gtcgtggcac ccctgctggg tgatgtgccc agggctttcc 1320

acgatgatcc cactgaatcc tcacatagcc ctgaggggaa gctgccacca tcatcacctc 1380

cacatcacag aggagtacac aggcaggaca gtcacactcg gacaagtggt ggagccagga 1440

cacaaaccca ggtctgtctg gactctaagc ttaggctctt accatgaact gtgatccaac 1500

catcgggaga gtcagcccaa tgaccttcaa gttccctcca atgctaaggt cacacctgag 1560

aagcttgcta gggtttctaa agcatcttcg tgcacatatg tccttccagg gttcttccca 1620

gcagtttgtg agatgggcag ggcagatgaa gggaaaaggc ttcaagacat taaggaaagg 1680

gcccaagatc acacccgagt caggatcaga gcaggaattg gaatctaatc tctacacaga 1740

gttgccagat accaatctgt gcttctcatg gttagaattt acctacttat ccaactattc 1800

acccatccaa ctacccaccc acacatgtac ccaccagtcc acgcattcat ctatccagcc 1860

atccactcac ccacccaact atccactcat ccattcacct accaatccac ccactcatcc 1920

acccatcaat cactcaccca tctatacatc catccaccta cccctttctc cctccctcct 1980

tctttccctc cctccctttc tttcctcatt cagactgttt attgagagcc ttgaaggtca 2040

ggcacttgga tggctgctgg ggagatgaag cttgaagatg tgtgaacggg ctgctcctct 2100

tggagacatc atcttttccc tcatcagtga gggttctgga aggggaatgt gggttgggag 2160

gcaggcagga tggagaggtg gctgttgagt tcctgccctt tgggttttgg gtcacagtac 2220

tagactcata gaattgggtt aaaaacttgt attttttttt ctaaggagaa gctggaaagt 2280

gaacaagtaa gatagggcgt ctggctcctt gctgtgtata gaacagatac ccagcaatga 2340

ccataggtgc agagtttatt tatgtagaga ctttttgaat ataagaaaag aaatgacaaa 2400

tatatatgag ttactatata atgaatataa gtataattaa aataaaccaa ctataggtag 2460

tgcagggtga gtttccaata ataaattcta aaccaacttc catcattcat ggaaccagat 2520

attctattca ggaattaata tgcattctcc catacatttc ctattataca catgggaggt 2580

agctactcat attaaatcaa tatcaaatag atgggaacat gaggagagag attagctacc 2640

aaggtaaatt gtcatctctc agtggcagag aaatttgtca accgaggtct tcctagaaat 2700

atgctctgac ctaccataca attcttcctg cctgaggaac tggtaaaaac tctccaagtc 2760

catcctggct tccttctttc cttcccaact tctgctcatc ctgacctgac cttgtcccaa 2820

gctgctctat ggcttttatt gctctttgca gaaatcgctt aaactttttg tttgcagttg 2880

ctccctcctc tgtgtttaac tgtattttaa gggggggttc attttccaat cctctttgct 2940

aaatgtcggc tgctaggtac tcatcccccc tggaatgccc tctctttcca tttttatgac 3000

tcagcaactt tccctgggct tcctgcctct gagctggctg gagctgtgag acaatttcgg 3060

gctttgggca gcacctaaga catacaggat gcgaatccat gttaggatcg tagtgacctc 3120

ttccatataa tcccccattg tactactcct caagagagaa cactctgtct tctcatcagt 3180

gtaatctcaa accctgcacc ctgactgttg tgggtgtagc agacacacaa gaaatatttg 3240

ctgaatgtta agtgaattgg aataaacatg ttgatttaag tgacaaaacc ttctattttt 3300

cataataata aatgtataat gattgtcaaa aaatgaaaaa agttagatgt acaaaatgaa 3360

aaccaatatt acctacagtt tacccccgca accggtagcc ccggggaata gatgtggata 3420

cggatgtgca tctaggtatg gataaaaatg cagggattca tgcacacaca ggtcacgtct 3480

tatctggtgg cctctttggg ataatatgtt caaggctttt cagatgccat tcgcatcatc 3540

gaatacagca tttttatagg atcttacatc ttaaattggc ttaatgtttt gttctccgat 3600

tataaaatct gaaaacagga aaaattaaat aagaatacaa agcaagtctc tgatcctgca 3660

ctgcccaaga ggccctgtta gcgcgttggt atggcgcctt ccagatcttt ttctgcatga 3720

aatcatgggc ttatcacaaa gatgggaact tctttcctca cttcttcttg ccctgttgtg 3780

aacatctgtc tatggccctt ctccaaggcc atctggggat gtggatgtga ggctggcatc 3840

tgctctcggg ctcttttctg tcacgttttg ctgttagctc cctggctaac atccacatgc 3900

actggctctc tggcatctcc atgataaatt cttatgagca ggatttctgg gtcaaaagag 3960

agtgcattcc caggctgtgg agacagggga cacgttcagg agggcctaaa aggctgaacc 4020

gtgtctcagc cttgaggcct ctcagaatcg ccagcttggg aagttaggag gcctgtggtt 4080

gctcatcact ccctcaccaa ctcgttctgc ccagggaagg tcctggggtg ggggcttcag 4140

agccctgtcc tggccacccc cctccctggg gtcctgctga caatcagggc cactgctggc 4200

tggttctgag atgccttctg gtggaccctg ggacccagga tggagctcag ggtgtgggga 4260

ggcagccagg gggcagcatc aggcccaggg gctggactag gagggggtgg ggtggggcgc 4320

aggctctact gcccccacct gtgagctgca caaacatcca gcagcagccc tgaaactgcc 4380

ccatgctcct cctgggccac acctgggcct gtttgtcact catcccatgc ccgggtggcc 4440

atgagctcag tttctcttcc tcttattttt ctccttttgt cactctgagt tctggtttca 4500

gccaacttgg ggttaaattt agcctgggga tttccagggg tggccagctg caggcagggc 4560

caccagagct gggaaagcgc atcccccaac cccatccctt ggcctggccc ccaccctact 4620

cccatctcag gcctccctgg ccaacccctt cacccaggaa tctgctggtt tcctgaatgg 4680

ctggcactcg gggtccctga cccagttcct ccttgaccag agggaaggct gggcccagga 4740

aggcaaaggc atttgcccag gttcacacag tgtcacccgc aggcccaggc aagagcaatg 4800

actgagggta ggtgagggtc cccataggcc aggcacagct taggcatcat ctcaatccgc 4860

aaacctacct tgtggggagg gactcttatc acccccttgg gcagatgagg aaactgaggg 4920

ctacagaaga gaagcagatt gctcaaggtc aagggttgat gagggctgag catggtgggg 4980

ggaccccttc tctgtctgac actgaggtct gggctcccag ggcagatgcc cctttgggaa 5040

gaacagtgct ctgggctccc acaggccttt gcagccaccc tccttccttg catctcccaa 5100

ggcacaggga aggaatggcc atcagggcct gtagccctaa ctctcatcca tgctgctttc 5160

aggcctcttt tctgcacccc tgaaatgggg ctggagctgt ggaatggtgg aggaagtgaa 5220

caatctggtc tctggcagag ggcagtggtc cacacagcca ccctccatac aggtcttctc 5280

catgcaggct cctccacacg ggtcccacca cacagcgccc tccacacggg cccctccaca 5340

caggtcccct ccacacaggc cccctccaca caggcaccct ccaacagatc ccctccacat 5400

gggcccctct acctggcacc ctaccgacag gcccctccat gcgggcccct tcacacatga 5460

tccctccacg cgggcccctc cacgcaggcc cctccacatg ggcccctcca cacatgattt 5520

ctccacacag gcccctccac atgggcccct ccacacatga tccctccaca cgggcccctc 5580

cacacatgat ccctccacac gggcccctcc acacatgggc ccctccacac gtgggcccct 5640

ccacatgcca ccctccatac aggtcccctc cacatgggcc cctctacatg ggcccctcca 5700

cacaggggcg cctatgtgag ctcgggcaga tccctttctc tcctggtctg tttctccatt 5760

tgtcagatgg gaggccctgg gctggaagtc aggacaccac agagctggct ctggcttttt 5820

gggatctaac tgctctgggc ctcagttttc ctacctgtga aatgggacca tgattctggc 5880

tgtggcctca cagaatccct gagtggctca ggtggataac atgtgtccac gtgttttata 5940

aatggtacgg gacgataagg ccactcaata gcgtctcaac acaggatgca gctacctgta 6000

gagttgcaga tgtggtcaga cctgaggttt agggcctcta ctctcggcct tgctctctct 6060

gggccctgcc cctgcccctc tgcagagctc tggggcttgg ggcactctgg tccactggcc 6120

tggaccagca tgaatgttca tggccttggt ccaccgtcca acccagagaa ctgtgcaccc 6180

tgcagggccc tgccttgggc ctggctccca gcatcctctg ctcattccct ctcctccacc 6240

gggtggtcac tacctgtgtc ctatgcaggg cccagacctc ttttctccct tgttccctgc 6300

ccccctggaa gcctggtcca tcttggagcc catccatgtg gcacagactc caaggacttc 6360

ctgggtatta agtagggtgg caggccaggt gacgtggggg acagagtatg gggcctttga 6420

ggtgagcttt cagggtgctt ccatgagggc ccttgtgcac tactgtggcc ctgtgggaag 6480

aagggatgct gtggtcactc ctataccttt gctcacactc acccctaaca cacccgtcct 6540

ctctctgccc ttcagtcctt gatccagctc ctaggcctgt ttgaggttgc tgctgccacc 6600

aggctacctt tgatgaccac atggaatccc cagtatctcc tgccccttgt cttcttcaag 6660

caagggggtc tcatctgcaa tctgagctct gggctgggcg gggacaagta cccccccttg 6720

cttgctttac ttcaggaact ggcctgggtg gctggcatcc tgtgcatgtt gaggggacgg 6780

tggtggggga tgccagtgga tgagctgatt gaagatggtg ctgggtctag ggccaccctg 6840

ttcagcctcg tgggtgggcc tgtggggagg aaggcagagg tgggggcact catccctggc 6900

ctgcccaccc tacagttcgc atcttatgtg tcaggtgaca tgtgcaatgg ctacgtgtgg 6960

cgggcctggc tcctggcttt cctgggccaa gggagctcca ggctggtgtc ctctaagaat 7020

gagctggtga agtcccacag gtggatggga tttgccgtct gcaggggtga acccaggccc 7080

tgaggctggg tacttggtct cctcccatac tctcatcaga acctggagcc ctggaatatc 7140

ctgggaacgt ttcctccaca tggagaaaga cagggaggtc tgtaacgggg gcagtgggta 7200

ggatctggtc atcggacagg gggcattaag cctcaccaca gtggcctgtt cctcacacct 7260

gtcaacctct ctttgacctc agcatctaga cccagcatgg ggaagactgg acagggaatg 7320

gggcacccca aggccacagc cccctcctcc catgagaagg cacactccag gtctcacctg 7380

ggggtgggct ggggcagtcc tgccttgcct gggacaccat gtgtcaagag ccctgcacag 7440

gccaggcctc ggggcttgtg gaaggatgag tgtgtgtgtg tgtgtgtgtg tgcgcgcgtg 7500

tgtgtgtgtg tgcatgttca gatatatgga tggacagaca cacaaataag tgactgacca 7560

gtctgaggtc ctgcccactg ggtagggacc ccggaagtcc tcttggcagc tgtgcttcca 7620

ggagcttcgt tactgcctca ggctctgggc tcccagggaa gctcttggaa gggttcttgt 7680

ttcccttccc atccccttac ttggcccagc tcccttgaag ggtccaagac ctgggggcag 7740

caggtggagt gtgatcagca gggtcccatg tccagctctg gccagccctg cccaccccag 7800

gcccctctga gctccaagtg tctggctgtg ccccccaatt gggcacagct ctgaacctgg 7860

catttagaac ggttcctggg gagctgtgag gagggggtag ggcctgcatg aatagaagct 7920

tctggagcag aaggtgctac ttctgatcat tgattgacca tgatctccag accctgtcta 7980

actcaagcct gggcacgggg cttgggtgtc ttggtgcggg gacacaggac cattctccag 8040

ctgggctggt ggaagctttg gaaactgaat cgcccccctg cctcccaccc tggggagaag 8100

gaggcacctg ctgccagggc tgaggcttgg tggggtttcc cagctatttc gggcctcagg 8160

cactcttggt ttcaccaggc cagctgcact ggggtgaggg ttccagggcc ggtggggagc 8220

ccagcatggg cagggggctg gcctgaggct gtctggggct ggaacagccc aaggatgaaa 8280

acaaccaacg gcagagacaa acaaaacatc tattttccat ctgattgtcc ctccccccac 8340

ccagagagtc agtttccatt attatcaacc cagctgcaca gaggggaaac tgaggctgag 8400

ctcgagccac taactccagc cacaggggcc tggcgctagt cagggctcct cagtcgctgg 8460

ctgcctccag cccctgggac gtcgcccccc gtgagacccc agccaggtgc tccctacccc 8520

ccatcctctg ccctatttct ccccagcaca aatcccgcct cctacatgac attgttcctt 8580

gtctgttgac ttgatttccc tctcccgcaa acatgagctg tctgaacctg gctgtgcctg 8640

ccttaaccct gctgggtttc caggagcagc aggtcctcag aagagagctg aacatgtgac 8700

ttcgctctgg gcagcccctg agaagggggc ttacttacct ggccggtctg gggtctcacc 8760

ttcttcagct actctgccca gcagcccctg gggaccagca ccccttcctt ctgggagctg 8820

ctctctgctg cccagacctt tgcaaacttt ctgtcccctg tgggagccca ggccaagtct 8880

gaccaccact tcccatcctc ctccccagac aggtcaggag aggatgtgtc taggggtctg 8940

agtgggaggc tcccctttgg cctgaaactg tgaagatgag gcccctttct cctctatcat 9000

ttccatagtc agaaagtccc aacatgtccc tgcacaggct gggaactggg tgggcaggga 9060

gggcccatcc tagtgacacc tgtgaggggt ccaggctttg gtgcctggca cggcaactgg 9120

cacattggat gctcagtgaa catcagacct gatgtgagaa gcaggaggga cccgtggcca 9180

gatcaagggt gttggtgttg ctataggaag agagagtcac catgaaggat ggcctgtatg 9240

gaaagaggca cactgcgggt gccctggtgg gtcgtcctgg ctgtgggcta ctctttcctg 9300

gagctgacac gggggctctg tgttgccagc cacagaagtc acccagcccc tccctggctc 9360

ctgtcccaaa gccaggaggc attctccctg ctgcctgtgg tctccccagg aactagggct 9420

gcccagagga gctgtccctt gccggtctct ggacgtggtg tgctgttgct gagtgggaga 9480

tggtattagg gccacccctg gttgtgttgt gagctgggcc tgagtgagct catttctagg 9540

ggctccaggc aggggaccag gaatgtcatg gtgactgcct ggatgtggcc cctagtgagg 9600

ctctcctggc ccctggcccc cattgtaatg tccagcactg cttaccatag tgctaggtgg 9660

ctgggcttgg cttcctcccc ctcattatta gccattatga agaacgtcat tcagtcattt 9720

agcaaacagt tactctgagc ttgggcaacc aaaatttgtt gtttaaactg ggatgttcct 9780

gggagtggaa ggggcactat taataacttc acgagtctct ctgatagatg tgggtggggg 9840

tcctggaggc agcccaggcc tgggtctctg ggcagagccc accattactg ctacaatggg 9900

ccaggcacgg ccaggcctgg gtcccgagct gaggctgggt acacataggc acctgggaaa 9960

ctgcagctga ttggtggcca gtggtctggg gacctggatg gggctaaggg tcaccagggg 10020

ctggacactg ctctagcatc cacctcagct tccagagtca gtggcacaaa ctgggaatgt 10080

cctggaaaat gcaggatgca ggtcatcccc ctcccctcag accttgccca gcccacgtgt 10140

ttcctgggga gaccaccctt gtgaaattac tggccaaaca gggtcaggtt tgatgtggtc 10200

ccaagtgacc aagtgacaaa gactgaccct gatcctgacc ctactgcccc tgctgatgtc 10260

actgtgttga catttccgtg gcccatctgc aatcctcaag atgaccatgt aggagagcag 10320

tttccccaga tcaccccaga aagagggagt cctcacgctt gggttgttac aagaggtacg 10380

ggtgcctttt gaggtgtttg cataactaag cattttggga aaagacctag tcttcccggc 10440

ctccgttgat gccagctagg gatgcttctg aatccccgag ggaactggag catctcacgt 10500

gtggcgcagg ggatcctggg cagatttggg gtcatttttc cttcacgtct cattcagcac 10560

agggaagagg accagtgtca agtcatgaca cacgactctt tcagggccgt agaagctggt 10620

ccttaggtca gggatggagc ccagggtgtg gggtcaacat caaggactcg tgtcctgcag 10680

gcggggtggg tggggaatcc agaacctggc cctggtgtga cagagactct ggagaagagg 10740

atgacatgtc gggagggagg agcaacatcc tcagcattgt gctgtgtcat tagcagtggg 10800

cattgcgagt ggatggtgat tatttcctct atgtccccaa aattcgcatg ttctagttgg 10860

ctggaaactt gattttggac ctttgtgttg aggaatgggg gcttgcctgg gatctggtgg 10920

cctcgttatg gggcatctga gctggtttgc ttgttcccag gtgagtgtcg atgtccccct 10980

tcttgcccct ggattcagtg aatatggggg ttgaagggta atggtgcctt tgtccaggcc 11040

tggctgggga cactctcttg gtggctttgc aggccccaca ggaggctggg cctgatggga 11100

gagtcaccca ctcagttcat agtagctccc agtcctttcc tcccgcaggg ccctgagcag 11160

ccacccccct ctcctccagc agggcccctg caccccctcc caggctattc tgggctctgg 11220

ctttgctgta gaggaagcca cagggcccct ccctgatctg actcagaaca cctccctcag 11280

cccaggctgc agctaagcag ggccagggcc aggacgagtt cctctcaccc catcccccac 11340

tgtgacactc ttccctatac cttgaggggt tcccccagca cagaccaacc ctccttcggt 11400

gtcatgaggc catcactcgt gaggggctgg ccccggtctg agctagacac agtggcatgt 11460

gaagacaggc gtgggctaag gcccaactcg gctactcctg aactgggtgg agcctgggca 11520

gcgctggtgc atccatcatg gttcacgtcc cctctgtgag gtgggcaatg gcagctttag 11580

gcctcagggt cactgtgagg gtcagtgagg gactgtgatg ttcccactgt ggcccaagga 11640

tcacagctgg ggctctctca gatggttctt tcagttaaat cttttcctga gtgtggggag 11700

atgctcctgc tgccccagac actccctggg cagagtgcca accttggaga cttgacaggg 11760

tgggggcagt ggtgtctcca tgtcgttctc caagcttgtg agctcctcta gggcaggtgg 11820

tcctgggctg catacaccag cccctggccc acagctgaga gtttgttgaa tgaatgaatg 11880

agtgagtctg aacaaatgaa tggatgaatg aatgaatgaa aagcccttag ggccgaggag 11940

tcatgaaaag aggcgggcat gggtcagccg gtaacctact gtgtgaccct aggtgggtca 12000

tcacccctcc catcggagcc tcagtcctct catctgtgaa atgggcacat gaacacctgt 12060

cctggctctg ctttgtagac tcagagggca cccccatggc tgccctcctg cctgcctctt 12120

tcacccactg agtcaacaga gaatgaggag cccaggtctg cccatcccac agagccacaa 12180

gggggttcat gcactcagca ccatctcttg gccttgccca ggcacggcct caggatggaa 12240

gggtggccta ggtcaggggc ctgggcaacc actgtgctgc cctgaacacc tgcaggacct 12300

gggcaggctc agcctcctct gcacgcctta gggaaggggc tggctctggg ttgaccttgg 12360

atcagagtcc cagtgcccac catctgtgca gccctgggca gctgtgggac tcgggcaggc 12420

tgctggtctc cgaatctccg tctcctcatc tgtaaggtgt gcgggacatg gacacgtgcc 12480

tcctggggtg gtttgggatc ccatgacata gagaggacag acgccagtgc agggtttccc 12540

tggaagtggc cctgggccag gtggcaggtg aggtagcatc ttcagcagag ccgggagcag 12600

ttccactgtc gagcagtggc catcgagggg ctcagtgata cccagcccag cacctggctg 12660

aacatcggag gacgtgaggg gtgcagcccc tgcttccagg ctcccgtggt gtcccagatc 12720

ggtgctgggc agggcggcac caggagagcg tggtcagcag ccctgggcag ccatcaagcc 12780

tgaggcacca ctgggctgag ggctggggcc ggggaaggag caaggaaccc ttttacattc 12840

cggtagttca tgaatcacac agtgaaagaa agagtcacca agggtgctag ctgccggtgg 12900

ccctgatggg gcagcagcag aacaaagtgg gaggggacca tagaagagag gaccttggcg 12960

gggggtgggg gtgggggtgg gtgggtgagg ggcttcaaga tgacgagaat gatgtgagtt 13020

gccaagccca tcctgaggtc aacagaagga ggacagtgag gggaccattg gattgagcca 13080

cgtgcaggcc actggggctg tgatggggtg gtggtgagac tgctgggctt gaggcaggaa 13140

gggaggggac cgtggaggcc aggagccctt gattaccact tgtaaaaggc aggagagagg 13200

gaggggcagc cagaggaagg cagcattcac attttattac ttcttcctgg caggaccaca 13260

tttgtgctcc taaggtgacc atcaggaggg tctctggccc ttcatactga gatgttgagg 13320

cagagagact gtggtccagg ttctctccag gaagagccca ccattactgc tgcagtgggc 13380

caggcaggac taggtctggg tcccaagccc aggctgggca catatgggtg cttgggaaac 13440

catagttgat tggtggccag tggcctagta cctggatggg gccggggtca ccatgggctg 13500

gtcactgctc cagccccagc atcaacccca gctcccagag tcagtgggga aggggactgg 13560

agtggggtcc gattacagtg accacaggga aatgggggcc acagggaaat gggtgcgggg 13620

agcaggggag attcgggagg tgggtggaag aggaccacgt gctcccatcc tgacagccca 13680

gagcctgcgt gtgcctgggg cggctgctct ctcaaagggc atgcgtctgt cttggggact 13740

gagggtctct gaagtcaggt cgcctcttgc tcgaaagccc cctgccacct tgctcttgac 13800

tttagcacct cctcacaggg cagcacgtgt tatgttatcc gggaggaaat ggaaaccctg 13860

ggaggttctc tggtggggga agggaggagc ggaggtagac taagaggggg tcagcagggg 13920

gagaaggggg aaccagccca gagttctcca gggacgggtg cgtgttgtcc tagcctgtgc 13980

cctgggagtg acaccaggtg agttgagatt tcagtaccca gcatttttgt gactgaagta 14040

aagaaaattc tgatttggcc cttgcctgct tgtacaagtt aactgcttgc tttttgccca 14100

gcatcccttg ttcacacaat gtgatcataa gctgcagatg tcctgtttct ttgtcaagaa 14160

ggaggagata acttcagagt cacaaaagta ttttgcagga gtgaatgcct ttagggacgt 14220

agagaccagc cgctgtcgcc cgcaagtcta gttgcttaag atgttacctg agactgaata 14280

aacacagact tttcccctga aatgccacct ccctcactcc ttagccacat aaaaactccc 14340

tgcttcatcc ttttgttaac atggagtgga ggaatttgct ttcttgtctt cttactttag 14400

ccaaatgaaa taaatctttt tctatctcca agcacttgtg tcagtttttg acctccgctg 14460

catatcaggt acacgaacct gaatctgggg ttctacaaag agtctcacat gtcaccactc 14520

agcccccctc gaacacctac tgcgctcggt gctgggcccc gagaggaccg gggttcctct 14580

ttcccacagc cccagaccca gagccccatg tgctcttccc ttgggagagc agtccctgac 14640

tgaagatgtc aggaattttt ggatgttgag ggagcaactg ggaccaggaa tggaggggat 14700

tgggcttggg gagccacctg ccagggactg actctcctgg aggtggggga caccgtaagc 14760

cctggatcca gaaccaggac tgagatgggg tctctaggaa aaggggactc tcatggtccc 14820

tccctccgag tgccctgacc ctgatatttg acccggagga aggagaaggg gcatgaggaa 14880

tctggctact ccaacctgag gccccgtccc cttccatccc ttgttggtcc aataattctg 14940

tcacttgaat cccgggtccc gggatggggg cctgggtcca gtgggctttg agctctccag 15000

ggaggaagct ggatgaagtg ggtgggcgtg gccaggtggg ctgagggcag ccaggcacct 15060

ggggtccaga gctgagatct gcagttgggt ggaccaggtt tgagtcccta ctgtgacctg 15120

tggcaggtca ctcaggctct ctgggctgtg atgtcaccat cctagagtga ggatcacaat 15180

aggatcttcc tccaagggtg atttggcctg gagggcattt agtctcaggg ggcctgctgg 15240

aagacagtgc cctagagctg tggctaaact cccagtgcct tcctggaacc ccaggaggag 15300

aaggggccac aagagaccag ggatagggag ggcttgggct gggctgggac ggccccccag 15360

gagtggctca gcctggactg ggcagggcat ggagaaaggg agaggaactc gggaaagggg 15420

ctgtggtcgg ctaaattagg ccccgcgggg acagtcgtgt cccaatcctg gacctgtcta 15480

tgtgtcatgt gtggccacag ggactttgca gatgtcatgt caattaggag cctgaggtgg 15540

gaaatgctcc tgggttatcc aggtgggtcc aagctggggt ctttgtagaa gaaagaggga 15600

ggcaggaggg tcagagtgag aggacaagat gtggggatgg agggagacgg agagagagag 15660

aaggaagaga gggagagaag aaggtaggga gggagagaag agcaagagag agatttgaag 15720

atgctgagct caggctctga agttggagga aggggccagg agctgaggtt gtcctctaga 15780

agctggaaaa gtcctggaaa cgaattctcc ccagagtccc cagaaggagc cggccctgct 15840

gggaccctga ttttagcccc caaggactca ttttggactt ctgacctcca gacctgggag 15900

agagtcagtg tgtgatgttt taaggcacta aattaggggt catttgtgat aacagccagc 15960

agccacagga aagtcaaaca ggagccgtgg gagagactcc ccaggatgcc acaagggccc 16020

ctcccaggcc ccacccacag taacccaggc ctgacccctg caggtgcccc cactctacag 16080

aggaggggcc atgcaggagg ggctgggtgg ggctgggtcg tgttctcttc ccccacccct 16140

tcctctcaga gagcagttgg agccccagac tcagggaacc tcacagagaa agtcctgatg 16200

agggacctgg ttttccagtg aaggcccggg atccaatgga gaagtgttcc caaaaatgat 16260

gttcccaaag acggccagtg actttctgtt cacgcacttg agagaatcac catgtctaag 16320

ggctgccaga tagttacctg tgcgtgtgtc aaggttcaca tcccattact actgaggcct 16380

tggatagaac atgttcccac ttttagttgt gacgagatga ctggccccag cattgagaac 16440

accaccttcc cttcagcaaa gggtttgttt aaatttacag aagtggtgaa gtgcttatta 16500

ccctccttgc actggttcct agggagctca gagcctatct tttccaagtg tcccgagcct 16560

cctgtaggcc tttcgtgtga gtctccttcc tggcttcctc ccattcctct taagtgtctg 16620

aacctgctgt caggtgagga atcgaggtgg gtccctcatt gaacagcttg agaagtcagt 16680

ggtgctggag tgtcctggac agtctggctc tcagggatga cagggagggg gtgagggtgg 16740

gtgggagctg agctgggcac agagggagga gggcattccg ggggcaaggg ccaggggcaa 16800

agaggctcca ggcttggagc agggttttca ggctgggaaa agagatggtg ggtgaagggt 16860

gtctctgtta gggcagagat agagggtgta actggagtcc cagccagaga caactccatc 16920

caggcaggag gcagggaggt ggggggctat gctttgagga gaagggggag gaatgaaagg 16980

gcagagggac tggtgttcag tgatcagact tctcctatcc gggcctcttt caccaggaga 17040

caccccggga ggctatgagc ctctccctca gggccacttt ctcactgggt gccaggattc 17100

ccccacccaa cactgagaac ccaggggagg cctcaaatta tgtgtgagat aagaaaagaa 17160

gagagggagc aggatttctc gttgacgatg aggatattta ttgagaaggg ctggatgact 17220

tgggatgggg gcagagaccc ctcccctggg aacctgcagc tccaggcccc tgtgggtggg 17280

gtgagggtcg ggggcctaag agcattctgc aggggccact gtcttctcca cggtgctccc 17340

ttcatgcgtg acccggcagc tgtagcttct gtgggacttc cactgctcgg gcgtcaggct 17400

caggtagctg ctggccgcat acttgttgtt gctttgtttg gagggtttgg tggtctccac 17460

tcccaccttg acggggctgc catctgcctt ccaggccact gtcacggctc ccgggttgaa 17520

gtcacttacg agacacacca gtgtggcctt gttggcttga agctcctcag aggagggtgg 17580

gaacagagtg accgaggggg cagccttggg ctgacctgcg gggtggatga ggggcagggg 17640

gtcagagtcc tggtgtccac ctggggagcc cctgacctca gtcttgaagt gggtatggga 17700

ctaaggccca gggcagggac ctcaggccgg acccttgctg gactgaggtc aggtcagcgg 17760

tgagtggaga cacttgctgt cagatgggag ttgcccccac tcttgggggt cacagacccc 17820

ctcttggatc tgtctctttt tcatattttc catcacctat agacccccac cggtgctggc 17880

tccacactgg atgggatgga gtccaggacc acatctcacc ctgggtattt gtctatcttg 17940

ggggtgtctg tctgaacacc tcctaccagt tccccttgtt ctcatctctt catgcagggc 18000

agctgttggt cctgggttca ggctcttctc tctcacaggg agtgggtttc ttacccacac 18060

atctccctgg tgccatcatt gcagccccca ccccacctcc ctgcagagac ccctctgggc 18120

tgcctggtga gacagggtgt cagtcctggg ggctggattc cagtagatca aaggcctcca 18180

tttgaggcat ttttttttta ctccctgagc tgaacgtgga gctttgtctg ttcgaaagag 18240

tctccatgtc gtgagaagtt atctggggga tctgactctc cagtgggttt tccctgacaa 18300

acaggcaccc gtcccagtca ccgtccctac cagaagtccc tgtcctgggc atctgctctg 18360

aggtcttcct ccagtggagg aattggtaag catccacgtt tagggcctga gttttctgag 18420

acccaccacc ctgtctgtgg tgtcctgcct gtcttggggg atggcgtgtg acccagcgct 18480

gtggggccac cttctcacgg aatattcttc ctctatcttt gtcctcctgc catttcctcc 18540

cccatctctc caagtcctca gtcaaaggcc tcctgtcact gtgtttaatg cagccccgta 18600

actgggaccc acagaggaca gaatcacaga agagggcagg agcccaggca aggaggggca 18660

aagattccag acctccagac aggcttagac cttagccttc gacagacagg aaagggaaaa 18720

actccccgtg tttaacttgg gatctcaaag aggagagaag cggggagact taccgaggac 18780

ggtcagctgg gtgcctcctc cgaacacagc acacagtgac acagccccac acacaaaccc 18840

ctcctgacac acccgacccg ttccccgctg cagcagcttg ggcgcccagg tccagtggcc 18900

ctgtcatgaa gcaggtctgg ccgataggag ggtgaggggg atggcccctc catgcatgga 18960

tgcacacttt actggaagtc agggatccct tcacccccag agggagaggg gagagggagg 19020

aggcatgtct gcacatcaca tctgggtccc ctgctactct ctgtcctgga ggaagccatg 19080

tgtcccggct cagctccttt ggagactgtc accatggaag aagtgagaat cctgtttatg 19140

ccacagggaa ggtggagctg gtcccctatg gagatccgag ttccctggga gggtcctgga 19200

tgcccagcac agcccctagt gggatgccca tgtggtaatg attctaaata gcacaggaga 19260

gtgtgtgtgt atgtatttgc atgcaggcat gtttgtttgt gaatacctgt gtgcctatga 19320

ggtgttcatg tgtgtttatg tttgtgaaca tgtatgtatg tgtgtttgtc tcatagccca 19380

ggccccccat cctgtgctac ctcctgggac aatcatcaac ccggttctta gatcccacat 19440

acagctctta aaaagaccaa ttggtttaca ggcacgtcta tggcaccact ctggaatctc 19500

atgattgact ttgacctccc cagccccact gaccatccgt ggataactgg acattaggta 19560

cttgaggtgt ggtcagtttt gggctggtta cttaggggtg agagagagta gagttgaccc 19620

ttatccccca ggggacttga gtataactgg gacactggtg acacatgtgg gaccaggggt 19680

acccagctgg cctgtggggt gagtgacctg ggtccagagg gtcagggtcc agtctgtgcc 19740

tgcttcagag atagatgtgg gggtagaggg cagggtcctg ggagtcagga acagggtggg 19800

cccctaagga gtaggggaga ccaagggccc atggactgga gaggctcctt ccgcctcaga 19860

aggagggacc ccttgagaga gggaggctgt ccaggggtga ggcctgatga tccgtggagg 19920

gtactcgttc ttctgggcca tggggaacca aggagcggtc gccccaggcc gctggaggtg 19980

gacatgtcca cacaaacaca gcacgtatgg gaaaggggca gtcctggggc ctgctgccca 20040

aggctctgtt ctggaatatt ctctgggcct ttctaggtga tggtggaact acccctcaaa 20100

aacaatccag acttcattcc cacaaggcta aaagaatttt gttcatcttg caggttggac 20160

caatttaggg tagaaaatgt ataaacagaa gcaaaacaca aaacccaggg tttctggttg 20220

actgaggata tttattgagg gtttattgag tgtaggaaga agggctggat gacttgggat 20280

ggggagagag accccctccc ctgggaacct gcagctccag gcccctgtgg gtggggtgag 20340

ggtcgggggc ctaagagcat tctgcagggg ccactgtctt ctccacggtg ctcccttcat 20400

gcgtgacctg gcaactgtag cttctgtggg acttccactg ctcaggcgtc aggctcaggt 20460

agctagctgc tggccgcgta cttgttgttg ctctgtttgg agggtgtggt ggtctccact 20520

cccgtgttga cggggctgcc atctgccttc caggccactt tcacagctcc cgggtagaag 20580

tcactgatca ggcacaccag tgtggccttg ttggcttgaa gctcctcaga ggagggcggg 20640

aacagagtga ccgatggggc agccttgggc tgacctgcgg ggtggatgag gggcaggggg 20700

tcagagtcct ggtgtccacc tggggagccc ctggcctcag ttttcaggtg ggtgaaggac 20760

taaggcccag gatagggacc tcaggctggg cccttgctgt actgaggtca acttggcggt 20820

ggcggggatg cttacctcct gatgggaaac tcctcagcat ctgaggactg aggagaatca 20880

ggcctcttct gctccagcct cctctccaga ttctcccctc cttatcagcc agctctcacc 20940

tcacctatgc tccccgacgc tggctgggtt cagccctggg ccctcgtctt tccttggggg 21000

acgtctctgt ctgaacatcc cctgccaggt ccctcctctg ggtctgttca cgagggcatc 21060

tgtgtcctgt ttttaggttc tcctcccttt tgacaagtga atttcttcct cacatatccc 21120

ccagcaccat cattgcagtc tctgccccac tttcccacag aggccccttg gtggggaggg 21180

gatgtcagtc ctgggagccg ggtcctggga gtccaggaga tggagggctt gcatgtgagg 21240

tatattttct gttctgagcc tggcgctgtg tctgctctat ggggtctccc tgtcgttggg 21300

agatctggct gtcctaggac aaatgtctgt cctgacaggt ccctgtcacc tgcgcccacc 21360

aggagtgcct ccctcagcat ctgctttggg ggtctctctt tttgggtggg atggggaggc 21420

tctaaccttg gactgagtgt cctgagaacc accctgtcca tggcagcctg cctgtcccgg 21480

gggaggagga gggacccagc accctggcgc cacctcacag gacattcttc ctccatcctt 21540

gtcccccttc atttccccaa gtccccagtc aaaggcctcc cttcttggtg tttaaagcag 21600

ccccatagtt gggacccaga accctgggaa gctgaacttt ttccagcatc cttcccctgt 21660

cccatggccc tggtgcccca gcctgagccc ctgctctgct ggtctttggg gactggagac 21720

tctgtgcgag ggagcagaac atcccagagc ccaggggaga atgccccctg agtgacagcg 21780

gcaggttcag gcctgagcag ggagcattaa gaggtcactg ggatcttccc atcatgcccc 21840

accaggtgcc taagaaggga caggctggag gaatttgggg ccaggaaaaa aggcaatggt 21900

ggcagacccc agagagattg agacctctac cctagagaga acagaaagga cagaaaagtc 21960

tccccagatc tcaccctaga cccaaaagaa tagaaaaggg gactcaccga ggacggtcac 22020

cttggtgcca ctgccgaaca cattacactg tgatataacc ctgcacacaa accctcctga 22080

tacacccgac ccaggccacc cccccccaac cccctgcagc tgctcacaag gcccagggtc 22140

cagccccctt ttctcatggg tgggtgagcc ataggtgggg tgaagggggt caccttctac 22200

tgacctgaat atcacttcct tgggggctta ggggttcatc cacctcagtc atgggaaggg 22260

ttgacaggtc tgtgtgtgga attgtctccc aggtaattgc atttctgagg agttgggtgg 22320

cctggatcag ctggtgtcag gaatgccagg gaggaagggg aggtgagcgg ctgggtgacg 22380

ccactgacat ggtgcagctg ggacctcttt gagaagttgg gaatggtggg gagggtcctt 22440

gatgcccccc acatcctcag cactggaggt tatgatggga tgggcttgga atgggaattg 22500

ggggttgtgg gaagaatgac gttgaccaca gggacacaga ccccactgca aggtctgagc 22560

tccagcatcc ttggcccagc tgtcctgaag gagcagctca cagatcactg cccaagtccg 22620

cctcctgaac ccatggggac agcttcctac ctataggcca tgtgtgctgg attagagctg 22680

caatggccca gaggagagct tctcaaggga ggtcataggc ctcactggca gtgtggtttc 22740

ctccaggagc tctacgtgtg ggggatctct gctgtccact tcagctcccc cggcttacac 22800

acacacccac atgtgtgtgc tctgcagtcc cctacatgca cgggtgaccc acacagcctg 22860

tgctctggag tgtcaccggc acggggggca cccacagtgg atgggcatgt gcagtgtggg 22920

gcctggacac acacagcctg gacctgcatc ctggtctggg cccatcacct gaagagctca 22980

cacctgggac acactcctgg caccatgcag tccccaacag tattccccac agcaagcgcc 23040

tcccctgact gctgagatgt ggcctgctgc tccctgatca gaatcccgtc cacatcttct 23100

cctgtctccc tcttggcctg gacagaaaac cctgccccac cctcccatat ccaggggcga 23160

ccccctagtc ccaaagcctg gagacagcag gtctcagctt tgctttgctc atgtcctgac 23220

ctgggcctga ctgcagcccc ttctaccttc ctgatcctcc cagaaggatg cccagcatgg 23280

acatgtgtgt ttatgtgcat tgtgtgtgtg tacacgtgtg tgtgtattat agctcagatc 23340

ccatgctgag ttcttcagaa gaagaatcag cgtcacccaa gctcctggca taaccccaga 23400

atctgacttt gaactcacca gtcaattgac aacttgtgca cagcaggaca ttaggcattc 23460

ataatggtgt cagtgaggag cagagccgac ccttgtcccc taggaatgtt gaatacagct 23520

ggggcatccg gtgacacatg tgggatgaga ggcacccaga gggggatgtg gggtgagtga 23580

ccagggtcca gggggtaagc atcccatcag tgattattat agggagggcc tgtgggggct 23640

gagggcaggg tccctgggat gacgggaaca gagtgtgccc atggagaagg ggagatgctc 23700

taggctttga tgaggggaaa tacaggtggg gtgaccacat ggccttggag ggggatggac 23760

aaacaccttg agagagggat ggcatctgga cagtgagggc cgaggacaac aaagaaggga 23820

gctgagatgt tctcccgggg ccatagggat gtaaggaggg tgtgtccccg ggcagccaga 23880

ggtggacgtg ctcacagggc agcaggaagc acaacagaga cgtgggtaag aatgaagatg 23940

tctggttcct tctgaagact atttccttgt tctggaacat tctatgggga taaggtgatg 24000

gaaaaagtat ggctcagaga aacacgaaga tggtcccagg ctgcattgtc acaagtccaa 24060

aggattctgt tgatctttgg agataaaaaa atttaggatg gaaagtggat aaacaaaaat 24120

gagcagacag catgttttct gattgacaag gaggacattt attgagggtt tattgggtgt 24180

cggcagaagg actggatgac ttggggtggg agggaacacc tctgccctgg gatcctgcag 24240

ctccaggccc ctgtgggtgg ggtgagggtc gggggcctaa gaacattctg catgggccat 24300

tgtcttctcc atggtacttt ctttgtgcgt gacctggcag ctgtagcttc tgtgggactt 24360

ccactgctcg ggcgtcaggc tcaggtagct gctggccgca tagttgttgt tgctctgtgt 24420

ggagggtgtg gttgtctcta tgcccttggt gactagggtg ccattttcct tctatggctc 24480

ctgggtagaa gtcatttatg agacacacca gcatggcctt gttggcttgg agctcctcag 24540

aggagggcgg gaacagagtg accaaggggg tggccttgag cggacctgca gaatggatga 24600

ggggcaggag gtcagagtcc tggtgtccac ctggggagcc cctgacctca gtgcatgagg 24660

agtgttcagg ccccccagta ccagtgcagg acctctcaga tgtggtccac actgtctcag 24720

aagggtgtga catcatgttc ctaaaagaac tctgtcagtc agaaaacctc tgagcttggc 24780

ccctgcccat cgcctgtttg gaaatcttcc aaggacttgg gcttttgtat agacagtgga 24840

ccttcaattg cctggcgctg cctcaccctg tcatcctgac gtgatccggc tctggctgtg 24900

ttctctgagc tctgtgagcc tcttgttaga ttcactctgg aaacttttct cacctgacac 24960

agctgctctg ggttggggga gagtgggtct ctcacgtaag agctcttgtt tgtacccaca 25020

tccgtctctc ctggatcttc tgcagtggag tcagggcctt tgctcaccca agtcccactg 25080

tctgtctgtc ctgggctctt tgtctgtgga gtctgtgtta gagtctctct gtgtttaggg 25140

ccaatctaca gtttttctgc cttgggatat gcctgtcggt gaactcaggg gttcctgctt 25200

ctaggggtgc ctcccacaga gacccatcag ccacctccct gaatgccatg gcctttcgtg 25260

acacttcacc tggctccttc atgtccagag ccttcccgtc cctcctgaca gggcttcatg 25320

ggtcccaggc cagctcaggg ctctctccct gggagcccac agaaacctgg agaggcccct 25380

gcacctccta ccttgttccc agcctggatc cagagtccca cagtccaggg gactgggccc 25440

tgagactctg agttttcatt cccctatctt ggctctgagg gtttcaggtc cttctaaagt 25500

ccaccaggtg gtcactcctt cgagcccctt gttccctctc cctcatgaga tggggcctca 25560

gtttcccttg cacaaagttc gttagacaga agcccaggtg ggggaggaca gaactgagca 25620

agttttaggg aagcagcctg gctctggcag acagacagac agatgagtct cagtaggtca 25680

tccagtctag tgccctacat ctgaggtcag aggtgaaggg gtcttggtga cccactgagg 25740

taagggtcta aacagggagg ggagaggtcc ttgaccaggg cctaggctcc agagagaaag 25800

catcaccaca ggtcaacaca agatacagaa aagaacaaaa gcagcagaaa atttcccagt 25860

accttgggga gaccagggaa ggagggggaa aagactcatc taggacggtc agctcggtcc 25920

cctcaccaaa cacccagtgc tgtgacacag gctcatacaa aaaccctctc tggacgcctc 25980

tacccccaac ccccgctgca gctgtgatgg aggaagctga ggccacagcc cctggtggca 26040

tctgctcatg agggagttca cacggctgcc acactcaccc ccacagagag aaggggacag 26100

ctgacatcct gtgaccagca ggatccctag aaatcctggt tcctggagga cagtggattt 26160

ggttttccca ccaaccatca tgcagccatg gggatgagtt tattggaggg cactgtgtca 26220

ctaggagtga actcccatgg tcagggataa atggtctccc tggtggagct gactcgacac 26280

tgactgcttg gtgacgccct ggggtgatgc agagagtgcc ctctccctcc aggaagaagc 26340

tggttaccct agagtgcggt gtcttgggaa gcttcccatg aacctatctg tgactccagg 26400

agcctcagcc cagccttccc tgggcttgtc ctagttggga tgggcctggc cagtgaggcc 26460

tggctggtcc ccactggaaa tgccagcccc atcctccagg gtaaaatgga aggatctcct 26520

gcacccctca acctggtctc attttggtgg gtgaggtggg gtagtggctg ccactgcctt 26580

attcaaatgt gagggacata aaggagctcc cgagtgtggt ctccaaatat caccttctgc 26640

taagggaggt gaggtcagtg cgggatccca ggcctggggc aggaaaggga catgatgaga 26700

cagaacctct catcctccga gaaatcagga gccatggaag ctcctggggc aatgggagaa 26760

aatcagatgc cacctgagga ggcgcccagt gggtaaatgt ttatgttttg tagccgcatt 26820

gtcattgcaa ttcagtatat ctgcagtgga gcccacatgt cagatgtcct cacacgctag 26880

agctcactcc ggtactacat tgtgctgtgc gccctggagg atggctgggc accagatcat 26940

cagttatcaa agtggtgaac aaaagggaag aatgtacatt ttttcttctt cccaaagcat 27000

gaagcacact cctgagcgca cggtgcagct gctgtgccct cccttctgga agccctcatg 27060

agaaggaggt ggaccgaagt cttgaccatc tgccttcctt ctcatctgag gcgcgaagtt 27120

attgatcatc aaatgcagct aatgtcactg aaagaatgac aacacaacat gggtggcttt 27180

gctggaagta gccacgtgtc ctgggaagcg agtttgccaa tgtccagcct gaaccccctt 27240

gagcctcttg atgctagaac catttttcag gaaaagagag gttgagggaa gcactgggtt 27300

cctgctcagg gaggtggcgc ctgcatcccg actgtgggag gtcccgggac cagctctctg 27360

ctcctgcagc tcctgccttg ccagggaaag ctagagcaat ttgggagaca cctgcagatg 27420

gaatgggtct aaaaacacat tcaccaatga taacgtcaag tattaatcca catagaaatg 27480

aaattaaatc acaaatatgc aacagcacca caaagtcgac aaaaatagac acaatgatat 27540

tcacaagaca tttgataaga ctctcattaa ctatttttga tgtcaagaaa tccttgatat 27600

tctctccagt tgtgtgtgtg atgactagca tttgcttcac acccttgctt gaaggagtag 27660

tttagttatt ctgctgtggg tgggtccgca ttatctgaaa atggatgatg ctgcggactg 27720

gctgatggga accttggagt cattactatt cttctgtctc cctatttttg tgttcagaga 27780

tcttcatcat aaaaatgtac aacttataga gagggtgaag cttgggacag tgtctcccat 27840

gggcttgaat tttcatagct gtgaaatggg tcaaacagag gctgagggac tctgggcacc 27900

tgaaattcaa ttcagctcac ggctgtgatt tggggcaggg gtggctggga ctgagctcca 27960

aaggggcccc tctctttgtg tatcattgtg gaaaaagccc ccaggccttg cagctctgtt 28020

cctgtcctgg ctggccatcc agcagctgct gccatgtccc tgatgaacca gaaacagtgg 28080

gggtcccagc ccgggcacta tggattaacc gagcctgtca aggccttcaa gggaccaaat 28140

tgcccactgt cagggacccc gggatgaggg tgtctctgag tgaggctgcc ggtgcaccat 28200

ggtgcagggg gtgaggggac ggaacaacgt gagtgacaga gggagtgggg cctggtggga 28260

tgggggtgtt tggggcagtt gagatcccgt gaaagcctga ctttgcctat agatagcccc 28320

taggtcatgt tcaccctgtg ctgaccttgc gtgacccagc cttgactcta gtccagaatg 28380

agactctgac acagactctc cccgacaacc atgggatgcc cgttctgtgc ctgggatggt 28440

gtctttgctc atttgtctca gtccacagga gtcctcccct cacacgagct gaccatcaca 28500

ggctcagcca cacaccaagc acatactcac tccaccccag agcctgactt tagaattggg 28560

aacttcatgg ttgagatctc agccacgtgc tggttaactt tgaacctcac cctaagccca 28620

tctgatcata actgatcacc cgtgtgcccc agcctcaccc cactttccag agctagcgac 28680

gtgaacacca gtatcttgca gagcatccta gggctcacta agttggctgg gatggtgagc 28740

aggattctgg acttcctgtt tccctattgc aggacttgta agtgtcttta cttagccacc 28800

ctgtattgtg aaaatctgtg aaattcacct gtgagtttcc attcccagac ctttccacaa 28860

gttcctggtg atggcaccac catggacttc tcacccgata aactgggcac ctggattttc 28920

ctcatccttg cttagaattt aaggaggttg caaactaaat atgagataag aaatgaagac 28980

agacagcagg atttctggtt gacaatgagg atatttattg agggttactg ggtgcaggga 29040

gaagggctgg atgacttgga gtgggagggg agacacatgc cctgatcctg cagctctagg 29100

cccccgtggg taggtgaggg tcggggacct aagaacattc tgcaggggcc actgtcttct 29160

ccacagtgtt cctttcatgc gtgacctggc agctgtagag gttgtgggac ttccactgtc 29220

gggtgccagt cttaggtagc tgctggccgc atacttgttg ctctgtttgg agggtgtggt 29280

ggtctccaca ccctgggtga tggtgatgcc atctatcttc caggccactg tcatggctct 29340

cgggtagaag tcactcatga gacacaccag tgtggccttg tcttggagct ccctgcctct 29400

ggggtcagag gagatgaagg tgtctcagtg accagctgag gcatacaggg gtctgaatag 29460

ggaggagaga gatcagcctc ccaggaccga ggtcccaaag gaaaccctga ccacaggctg 29520

tggcaagtca cagacaaaga tcgaaatcag aataaaattg tcaccaactt ggcagggaaa 29580

gaaagggaag aagagactca tctaaaatga tcagctgggt tcctccacca aatacaaaat 29640

gcggtgacac aggttcatac aaaaactctt ccctggggcc cctaactccc cctccccatc 29700

ccccacctga agctatggtg caggaagcca gaccactgcc cctggaaggc atttgctcag 29760

cagagtccac acacctgcct gcccttctca agctgagatg ggacagtcga ccaagcgtat 29820

gcagtaacgg gacgtctgga aatggcgagg cctgaggagg ttggtgtctg gatcaggttt 29880

tccaggtgac tgtgacccag gaatgggaat gagctccttg agggcatttg cttcacgaag 29940

gagtgaagtg acatggaagg ggacagggat tctctatagg gttgttgtaa ccctgatctc 30000

ttggtgtgac tacagggtaa gagcaggagg cgtcctcagg tgtccccagt gagtgcctta 30060

gaggaggggc agcttgcccc agagtcagca gtcatgggaa gctccaacgt gaccacaacc 30120

acaactccag ggcgtctcac ttccgtcctc cttgtgttgt cttagtttgg gatgggtctc 30180

actaggtttt agtatgaaat gccagcctca acctccaggg agaagttgga agcaagcccc 30240

tgttaccctt tcagggtctt gtctcatttc tgggtatgtc tggagtgtct gactcaaaag 30300

ggctgtgtga gaagcacccc aagtgtggtc actaaatccc acctcccagt tatggggact 30360

agtccacttg gggttaatgg gcaattccag gcacaaggca ggagaaagat gtgcccagga 30420

tagattgtga acagagagcg gaaagccatc gaggtctcct gggttccttc aggaggatcc 30480

tggggccacc agaagaggca ctgagtgggc cacggtgggt cattgtcatc atcaatgaaa 30540

tagaaccatg gtggactcaa gtgagtcaca tacagtaaca taactttgag ctcaccatga 30600

aataattgtt catcttcacg agatggcagg caacatggtc atcattatta agatgctaaa 30660

tgaaaggaaa gactcaacat gtctatgcta tttcttactg aaataagaat cacactccca 30720

gaggagcaaa gaacatatga gggcacacct tttacagaca gtcagtagaa ggaaaaggtg 30780

gtcgaactga aatgcagtca ccttgcatcc atataccatc taatggcact ggatacaaag 30840

tgtgaagccc agctaacgtc acaaatagat gaccccacat gggagcccag tagccaagcg 30900

gtcttgctga aaagtcacac tgagccttgt taaacctttc tggccaagaa ccaattttca 30960

ggaaacgcag aagcagagag gagcgcttgt ttccagcttg gggaggccac aggctgtatc 31020

ctgaccatgg gaaggtcctg ggaccaaccc ccagctcctg tagctcctgc cttgcaagga 31080

aattaaaaat caatgaggaa gatacctgca gacagaacag gtctaaaaac acagtcacca 31140

gtcacaattt caaaccatat tgaatcctca tagaaatgaa atggaaaacc acagaataac 31200

aacaacacaa acacaggtct agtgaaagaa gcataggata tttgtgaagt atttgatggg 31260

aaacattctt tttttgatat gaaagaatta gatatttata tttaggtttg taagaatgta 31320

gttttttggg gggggtccat atttttagag acacaaatgg aatatttaga ctaatggaca 31380

tgatgtctac aatttgcttc aaacaactgt gtaaaagagg ttacttgatt gtgggtgggt 31440

caacatgagc tgtattatcc tgttttttta ttttcttgct gtcagtttcc gtaatatctt 31500

acaaggcagt aattagagag attagtcttg ggacagcacc aggggacatc cagaggggat 31560

cctacccagc agggccattg agaaggtgga gatcaaagcg gtcctctttg ctataggggt 31620

ctcagcgttc acatctatga aatggggcca gacagggtgg tgggaatcag gacccttaag 31680

tttccttcta acttgtaggt tgtgacttga ggcaggggtt ctatgattag gctccaagag 31740

gaaccccttc tctgccccac tatggcccta gagttctctg cccttgcagg ctctgtctca 31800

ggcctggcct ggccacccag cagctctggt gaaggcacct gccacaccgc tgatgtgctg 31860

gaggcaccaa gaaggggcct gacagtgccc aagactgatg aggagggctg ggaagtgcat 31920

ctcccaaggc cacctcactc gctgtcaggg accctgggat gattgtgtct ctgacatcag 31980

ggtgtaggtg cacatggagg ggctgaacca catgattgag ggagggagta ggccctggtg 32040

gggaggggtc taaggtagct gagtcccatg aaaacttgac ctgcctgtag cccttaagtc 32100

acacttaccc tgccctgacc tagtgtgacc tggccctgat tcttgggcga gtatgatttt 32160

gacacagcct ccatcagaca accaatggat gccgattcta tgcttgggat ggtgcctttc 32220

tcatttgacc 32230

<210> 83

<211> 106

<212> PRT

<213> Intelligent people

<400> 83

Gly Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser

1 5 10 15

Glu Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Val Ser Asp

20 25 30

Phe Asn Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Gly Ser Pro

35 40 45

Val Lys Val Gly Val Glu Thr Thr Lys Pro Ser Lys Gln Ser Asn Asn

50 55 60

Lys Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys

65 70 75 80

Ser His Arg Ser Tyr Ser Cys Arg Val Thr His Glu Gly Ser Thr Val

85 90 95

Glu Lys Thr Val Ala Pro Ala Glu Cys Ser

100 105

Claims

1. A method of identifying an individual having non-small cell lung cancer (NSCLC) who is likely to benefit from treatment with an anti-PD-L1 antagonist antibody, the method comprising determining the expression levels of genes CD79A, CD19, BANK1, jcha in, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual, wherein an immune score expression level of the genes that is higher than a reference immune score expression level of the genes identifies the individual as an individual who is likely to benefit from treatment with an anti-PD-L1 antagonist antibody, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

2. A method of selecting a therapy for an individual having NSCLC, said method comprising determining the expression levels of genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from said individual, wherein an immune score expression level of said genes that is higher than a reference immune score expression level of said genes identifies said individual as an individual likely to benefit from treatment comprising an anti-PD-L1 antagonist antibody, wherein said anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

3. A method of treating an individual having NSCLC, the method comprising:

(a) Determining the expression level of genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from said individual, wherein the determination of the level of immune score expression of said genes in said sample is greater than the reference level of immune score expression of said genes, thereby identifying said individual as one likely to benefit from treatment comprising an anti-PD-L1 antagonist antibody; and

(b) Administering to the individual an effective amount of an anti-PD-L1 antagonist antibody,

wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

4. A method of treating NSCLC in an individual who has been determined to have an immune score expression level of genes CD79A, CD19, BANK1, jchanin, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from the individual that is greater than a reference immune score expression level for the genes, and who has thus been identified as an individual who is likely to benefit from treatment comprising an anti-PD-L1 antagonist antibody, comprising administering to the individual an effective amount of an anti-PD-L1 antagonist antibody, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

5. A method of identifying an individual having NSCLC who is likely to benefit from treatment with an anti-PD-L1 antagonist antibody, the method comprising determining the expression levels of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from the individual, wherein an immune score expression level of the genes that is higher than a reference immune score expression level of the genes identifies the individual as an individual who is likely to benefit from treatment with an anti-PD-L1 antagonist antibody, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

6. A method of selecting a therapy for an individual having NSCLC, the method comprising determining the expression level of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7 and IGLL5 in a sample from the individual, wherein an immune score expression level of the genes that is higher than a reference immune score expression level of the genes identifies the individual as an individual likely to benefit from treatment comprising an anti-PD-L1 antagonist antibody, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

7. A method of treating an individual having NSCLC, the method comprising:

(a) Determining in a sample from the individual the expression levels of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5, wherein the determination of the immune score expression level of said genes in said sample is higher than a reference immune score expression level for said genes, thereby identifying the individual as one likely to benefit from treatment comprising an anti-PD-L1 antagonist antibody; and

8. A method of treating NSCLC in an individual who has been determined to have an immune score expression level of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from the individual that is higher than a reference immune score expression level for the genes, and who has thus been identified as potentially benefiting from treatment comprising an anti-PD-L1 antagonist antibody, comprising administering to the individual an effective amount of an anti-PD-L1 antagonist antibody, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

9. A method of identifying an individual having NSCLC who is likely to benefit from treatment comprising an anti-PD-L1 antagonist antibody, the method comprising determining the expression level of the genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual, wherein an immune score expression level of the genes that is higher than a reference immune score expression level of the genes identifies the individual as an individual who is likely to benefit from treatment comprising an anti-PD-L1 antagonist antibody, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

10. A method of selecting a therapy for an individual having NSCLC, the method comprising determining the expression levels of genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual, wherein an immune score expression level of the genes that is higher than a reference immune score expression level of the genes identifies the individual as an individual who is likely to benefit from treatment comprising an anti-PD-L1 antagonist antibody, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

11. A method of treating an individual having NSCLC, the method comprising:

(a) Determining an expression level of genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual, wherein determining an immune score expression level of the genes in the sample is higher than a reference immune score expression level of the genes, thereby identifying the individual as an individual who is likely to benefit from treatment comprising an anti-PD-L1 antagonist antibody; and

12. A method of treating NSCLC in an individual who has been determined to have an immune score expression level of genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual that is higher than a reference immune score expression level for the genes, and who has thus been identified as an individual who is likely to benefit from treatment comprising an anti-PD-L1 antagonist antibody, comprising administering to the individual an effective amount of an anti-PD-L1 antagonist antibody, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

13. The method of any one of claims 1, 2, 5, 6, 9, and 10, wherein the immune score expression level of the gene in the sample is greater than the reference immune score expression level, and the method further comprises administering to the individual an effective amount of the anti-PD-L1 antagonist antibody.

14. The method of any one of claims 1-13, wherein the reference immune score expression level is an immune score expression level of a gene in a reference population.

15. The method of claim 14, wherein the reference population is a population of individuals having the NSCLC.

16. The method of claim 15, wherein the population of individuals comprises a first subset of individuals who have received treatment with a PD-L1 axis binding antagonist and a second subset of individuals who have received treatment with a therapy that does not contain a PD-L1 axis binding antagonist.

17. The method of claim 16, wherein the reference immune score expression level visibly distinguishes each of the first subset of individuals from the second subset of individuals based on a significant difference between the individual's responsiveness to treatment with the PD-L1 axis binding antagonist and the individual's responsiveness to treatment with the therapy without the PD-L1 axis binding antagonist that is greater than the reference immune score expression level, wherein the individual's responsiveness to treatment with the PD-L1 axis binding antagonist is significantly improved relative to the individual's responsiveness to treatment with the therapy without the PD-L1 axis binding antagonist.

18. The method of claim 16 or 17, wherein the therapy without a PD-L1 axis binding antagonist comprises an anti-tumor agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, or a combination thereof.

19. The method of claim 18, wherein the therapy without a PD-L1 axis binding antagonist comprises a chemotherapeutic agent.

20. The method of claim 19, wherein the chemotherapeutic agent is docetaxel.

21. The method of any one of claims 17 to 20, wherein responsiveness to treatment comprises prolongation of OS, prolongation of Progression Free Survival (PFS), or an increase in determined best overall response (BCOR).

22. The method of claim 21, wherein responsiveness to treatment comprises prolongation of OS.

23. The method of any one of claims 14-22, wherein the reference immune score expression level is the median of the expression levels of each of the genes in the reference population.

24. The method of any one of claims 1, 2, 5, 6, 8, 10, and 13-23, wherein the benefit comprises a prolongation of OS in the individual as compared to treatment without the anti-PD-L1 antagonist antibody.

25. A method of identifying an individual having NSCLC who is likely to benefit from treatment comprising an anti-PD-L1 antagonist antibody, the method comprising determining the presence of a Tertiary Lymphoid Structure (TLS) in a tumor sample from the individual, wherein the presence of TLS in the tumor sample identifies the individual as an individual who is likely to benefit from treatment comprising an anti-PD-L1 antagonist antibody, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

26. A method of selecting a therapy for an individual having NSCLC, the method comprising determining the presence of TLS in a tumor sample from the individual, wherein the presence of TLS in the tumor sample identifies the individual as an individual who is likely to benefit from treatment comprising an anti-PD-L1 antagonist antibody, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

27. A method of treating an individual having NSCLC, the method comprising:

(a) Determining the presence of TLS in a tumor sample from the individual, thereby identifying the individual as an individual likely to benefit from treatment comprising an anti-PD-L1 antagonist antibody; and

28. A method of treating NSCLC in an individual who has been determined to have TLS present in a tumor sample from the individual and who has thus been identified as an individual who is likely to benefit from treatment comprising an anti-PD-L1 antagonist antibody, comprising administering to the individual an effective amount of an anti-PD-L1 antagonist antibody, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

29. The method of any one of claims 25 to 28, wherein the presence of TLS is determined by histological staining, immunohistochemistry (IHC), immunofluorescence, or gene expression analysis.

30. The method of claim 29, wherein the histological staining comprises hematoxylin and eosin (H & E) staining.

31. The method of claim 29, wherein the IHC or immunofluorescence comprises detecting CD62L, L-selectin, CD40, or CD8.

32. The method of claim 31, wherein the CD62L or L-selectin is detected using a MECA-79 antibody.

33. The method of claim 29, wherein the gene expression analysis comprises determining the expression level of a TLS gene signature in the tumor sample.

34. The method of any one of claims 1-24, wherein the expression level of the gene is a nucleic acid expression level.

35. The method of claim 34, wherein the nucleic acid expression level is an mRNA expression level.

36. The method of any one of claims 1-24, wherein the expression level of the gene is a protein expression level.

37. The method of any one of claims 1-24 and 34-36, wherein the expression level of the gene is detected in tumor cells, tumor infiltrating immune cells, stromal cells, paracancerous normal tissue (NAT) cells, or a combination thereof.

38. A method of identifying an individual having NSCLC who is likely to benefit from treatment with an anti-PD-L1 antagonist antibody, the method comprising determining the number of B cells in a tumor sample from the individual, wherein a number of B cells in the tumor sample that is higher than a reference number of B cells identifies the individual as an individual who is likely to benefit from treatment with an anti-PD-L1 antagonist antibody, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

39. A method of selecting a therapy for an individual having NSCLC, the method comprising determining the number of B cells in a tumor sample from the individual, wherein a number of B cells in the tumor sample that is higher than a reference number of B cells identifies the individual as an individual who is likely to benefit from treatment comprising an anti-PD-L1 antagonist antibody, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

40. The method of claim 38 or 39, wherein the number of B cells in the tumor sample is greater than the reference number of B cells, and the method further comprises administering to the individual an effective amount of an anti-PD-L1 antagonist antibody.

41. A method of treating an individual having NSCLC, the method comprising:

(a) Determining that the number of B cells in a tumor sample from the individual is greater than a reference number of B cells, thereby identifying the individual as an individual who is likely to benefit from treatment comprising an anti-PD-L1 antagonist antibody; and

42. A method of treating NSCLC in an individual who has been determined to have a number of B cells in a tumor sample from the individual that is higher than a reference number of B cells, and who has thus been identified as an individual who is likely to benefit from treatment comprising an anti-PD-L1 antagonist antibody, comprising administering to the individual an effective amount of an anti-PD-L1 antagonist antibody, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

43. The method of any one of claims 38-42, wherein the B cells comprise CD79+ B cells, igG + B cells, and/or plasma cells.

44. The method of claim 43, wherein the B cells comprise plasma cells.

45. A method of identifying an individual having NSCLC who is likely to benefit from treatment with an anti-PD-L1 antagonist antibody, the method comprising determining whether the individual has clonally expanded B cells in a tumor sample from the individual, wherein the clonally expanded B cells in the sample identify the individual as an individual who is likely to benefit from treatment with an anti-PD-L1 antagonist antibody, wherein the anti-PD-L1 antagonist antibody comprises: heavy chain comprising SEQ ID

HVR-H1 sequence of NO. 1, HVR-H2 sequence of SEQ ID NO. 2 and SEQ ID

HVR-H3 sequence of NO 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

46. A method of selecting a therapy for an individual having NSCLC, the method comprising determining whether the individual has clonally expanded B cells in a tumor sample from the individual, wherein the clonally expanded B cells in the sample identify the individual as an individual likely to benefit from treatment comprising an anti-PD-L1 antagonist antibody, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

47. A method of treating an individual having NSCLC, the method comprising:

(a) Determining that the individual has clonally expanded B cells in a tumor sample from the individual, thereby identifying the individual as an individual likely to benefit from treatment comprising an anti-PD-L1 antagonist antibody; and

48. A method of treating NSCLC in an individual who has been determined to have clonally expanded B cells in a tumor sample from the individual and who has thus been identified as an individual who is likely to benefit from treatment comprising an anti-PD-L1 antagonist antibody, the method comprising administering to the individual an effective amount of an anti-PD-L1 antagonist antibody, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

49. The method of any one of claims 45-48, wherein the clonally expanded B cells are clonally expanded plasma cells.

50. The method of any one of claims 1 to 24 and 34 to 37, wherein the sample is a tissue sample, a cell sample, a whole blood sample, a plasma sample, a serum sample, or a combination thereof.

51. The method of claim 50, wherein the tissue sample is a tumor tissue sample.

52. The method of any one of claims 25-33 and 38-49, wherein the tumor sample is a tumor tissue sample.

53. The method of claim 51 or 52, wherein the tumor tissue sample comprises tumor cells, tumor infiltrating immune cells, stromal cells, NAT cells, or a combination thereof.

54. The method of any one of claims 51-53, wherein the tumor tissue sample is a formalin-fixed and paraffin-embedded (FFPE) sample, an archived sample, a fresh sample, or a frozen sample.

55. The method of any one of claims 1 to 54, wherein the NSCLC is non-squamous NSCLC.

56. The method of any one of claims 1 to 54, wherein the NSCLC is squamous NSCLC.

57. The method of any one of claims 1 to 56, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain variable region comprising the amino acid sequence of SEQ ID NO 7; and

a light chain variable region comprising the amino acid sequence of SEQ ID NO 8.

58. The method of claim 57, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO 9; and a light chain comprising the amino acid sequence of SEQ ID NO 10.

59. The method of claim 58, wherein the anti-PD-L1 antagonist antibody is atlizumab.

60. The method of any one of claims 1-59, wherein the subject has not been treated for the NSCLC.

61. The method of claim 60, wherein the subject has not been administered a PD-L1 axis binding antagonist.

62. The method of claim 60 or 61, wherein the individual does not have EGFR or ALK genomic tumor aberrations.

63. The method of any one of claims 1-59, wherein the subject has been previously treated for the NSCLC.

64. The method of claim 63, wherein the subject has been previously treated for the NSCLC by administering to the subject a platinum-containing chemotherapeutic agent, and wherein the subject is non-responsive to the chemotherapeutic agent.

65. The method of any one of claims 3, 4, 7, 8, 11-24, 27-37, 40-44, and 47-64, wherein the anti-PD-L1 antagonist antibody is administered as a monotherapy.

66. The method of any one of claims 3, 4, 7, 8, 11-24, 27-37, 40-44, and 47-65, wherein the method further comprises administering an effective amount of one or more additional therapeutic agents.

67. The method of claim 66, wherein the one or more additional therapeutic agents comprise an anti-tumor agent, a chemotherapeutic agent, a growth inhibitory agent, an anti-angiogenic agent, radiation therapy, a cytotoxic agent, an immunomodulatory agent, or a combination thereof.

68. The method of any one of claims 1-67, wherein the individual is a human.

69. A kit for identifying an individual having NSCLC who is likely to benefit from treatment with an anti-PD-L1 antagonist antibody, said kit comprising reagents for determining the expression levels of genes CD79A, CD19, BANK1, JCHAIN, SLAMF7, BTK, TNFRSF17, IGJ, IGLL5, RBPJ, and MZB1 in a sample from said individual, wherein an immune score expression level of said genes that is higher than a reference immune score expression level of said genes identifies said individual as an individual who is likely to benefit from treatment with an anti-PD-L1 antagonist antibody, wherein said anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

70. A kit for identifying an individual having NSCLC who is likely to benefit from treatment with an anti-PD-L1 antagonist antibody, the kit comprising reagents for determining the expression levels of genes MZB1, DERL3, JSRP1, TNFRSF17, SLAMF7, IGHG2, IGHGP, IGLV3-1, IGLV6-57, IGHA2, IGKV4-1, IGKV1-12, IGLC7, and IGLL5 in a sample from the individual, wherein an immune score expression level of the genes that is higher than a reference immune score expression level of the genes identifies the individual as an individual who is likely to benefit from treatment with an anti-PD-L1 antagonist antibody, wherein the benefit comprises an increase in the Overall Survival (OS) of the individual as compared to treatment without the anti-PD-L1 antagonist antibody, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

71. A kit for identifying an individual having NSCLC who is likely to benefit from treatment comprising an anti-PD-L1 antagonist antibody, the kit comprising reagents for determining the presence of a Tertiary Lymphoid Structure (TLS) in a tumor sample from the individual, wherein the presence of TLS in the tumor sample identifies the individual as an individual who is likely to benefit from treatment comprising an anti-PD-L1 antagonist antibody, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

72. A kit for identifying an individual having NSCLC who may benefit from treatment comprising an anti-PD-L1 antagonist antibody, the kit comprising reagents for determining the expression level of genes CCL2, CCL3, CCL4, CCL5, CCL8, CCL18, CCL19, CCL21, CXCL9, CXCL10, CXCL11, and CXCL13 in a sample from the individual, wherein an immune score expression level of the genes that is higher than a reference immune score expression level of the genes identifies the individual as an individual who may benefit from treatment comprising an anti-PD-L1 antagonist antibody, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

73. A kit for identifying an individual having NSCLC who is likely to benefit from treatment with an anti-PD-L1 antagonist antibody, the kit comprising reagents for determining the number of B cells in a tumor sample from the individual, wherein a number of B cells in the tumor sample that is higher than a reference number of B cells identifies the individual as an individual who is likely to benefit from treatment with an anti-PD-L1 antagonist antibody, optionally wherein the B cells comprise plasma cells, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

74. A kit for identifying an individual having NSCLC who is likely to benefit from treatment with an anti-PD-L1 antagonist antibody, the kit comprising reagents for determining whether the individual has clonally expanded B cells in a tumor sample from the individual, wherein the clonally expanded B cells in the sample identify the individual as an individual who is likely to benefit from treatment with an anti-PD-L1 antagonist antibody, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the HVR-H1 sequence of SEQ ID NO. 1, the HVR-H2 sequence of SEQ ID NO. 2, and the HVR-H3 sequence of SEQ ID NO. 3; and a light chain comprising the HVR-L1 sequence of SEQ ID NO. 4, the HVR-L2 sequence of SEQ ID NO. 5, and the HVR-L3 sequence of SEQ ID NO. 6.

75. The kit of any one of claims 69 to 74, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain variable region comprising the amino acid sequence of SEQ ID NO 7; and a light chain variable region comprising the amino acid sequence of SEQ ID NO 8.

76. The kit of claim 75, wherein the anti-PD-L1 antagonist antibody comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO 9; and a light chain comprising the amino acid sequence of SEQ ID NO 10.

77. The kit of claim 76, wherein the anti-PD-L1 antagonist antibody is atuzumab.